<%BANNER%>

Effects of the Timing of Initiation of Fat Supplementation on Productive and Reproductive Responses of Periparturient Da...

xml version 1.0 encoding UTF-8
REPORT xmlns http:www.fcla.edudlsmddaitss xmlns:xsi http:www.w3.org2001XMLSchema-instance xsi:schemaLocation http:www.fcla.edudlsmddaitssdaitssReport.xsd
INGEST IEID E20101130_AAAACW INGEST_TIME 2010-11-30T16:48:30Z PACKAGE UFE0010490_00001
AGREEMENT_INFO ACCOUNT UF PROJECT UFDC
FILES
FILE SIZE 5654 DFID F20101130_AABXSK ORIGIN DEPOSITOR PATH fickett_f_Page_017thm.jpg GLOBAL false PRESERVATION BIT MESSAGE_DIGEST ALGORITHM MD5
5bb10a4e9d03a093f8912095518823a0
SHA-1
c1397f2863ade4026174621a6fe2a0a51d6eb126
17677 F20101130_AABXRW fickett_f_Page_005.QC.jpg
1e0599a47a31b75ffe2d9b325bdbe7df
f8909f2073b83203f50326777f3f27e9dbefd834
1053954 F20101130_AABWPI fickett_f_Page_014.tif
8aab9f7875ecb79287b224eeaa3d9ca6
cb6d980a8c89f7162470297fba011a1a0f12db7c
51111 F20101130_AABWOU fickett_f_Page_018.pro
70aced9b123a566a3669e8d07b702fac
e46e6ea60046c74a6b546cd4388508b40eb16432
23801 F20101130_AABXSL fickett_f_Page_018.QC.jpg
0850d5217931ae63c323017a337abbdf
a63a6de8b03fb936c9c4dbbe064393e2181a6bc1
2810 F20101130_AABXRX fickett_f_Page_006thm.jpg
0f72ffdd5364c3f8a5036fa0cced0909
c1cbefc36d9f633948982576e86ba6b4de449299
23797 F20101130_AABWPJ fickett_f_Page_087.QC.jpg
b12abea12a0255314e26c35d842f7489
666ba4e2d1ce3483ccade41e1d0b7038b5f43e01
1262 F20101130_AABWOV fickett_f_Page_112.txt
1f4fec476d3fd58d3bc30ef9f422b3fa
76c5362267634967c8c484f4f4794af12930cc1a
6407 F20101130_AABXTA fickett_f_Page_033thm.jpg
cd1311c34d40395932b0c8f50f225a78
2ed97d5fe8effc3771379af2b6145013380e8589
6439 F20101130_AABXSM fickett_f_Page_018thm.jpg
34e49d387626bb32e23193f0a1c3323d
e9ce8c3eeda2043b8d912b8a5272eb8643bd0017
23856 F20101130_AABXRY fickett_f_Page_007.QC.jpg
d2af038486544708dfdbb127b10a9e1b
c2d8ba43e1310229b3cebd7276e965ac97490f46
2309 F20101130_AABWPK fickett_f_Page_056thm.jpg
42e2353975c388469693802c3c0eddf9
19acdc0430d23b552ddca12fdc438fe667d70dd0
889 F20101130_AABWOW fickett_f_Page_104.txt
337ebcb720861b93506b80812c5535c0
f5b6913980405bd5834e175b03f8ee3b87863603
24392 F20101130_AABXTB fickett_f_Page_034.QC.jpg
40376def6332da6a20d4de5e36d814e2
0a558c06fef5634173198702fcb5e657d7b38ea9
23792 F20101130_AABXSN fickett_f_Page_019.QC.jpg
79a1c9946dbe6a0d5bcb7d3a611010ea
cf46bb19e89e3d77e3cd5dddaa5436189764500d
12887 F20101130_AABXRZ fickett_f_Page_010.QC.jpg
d8e1da49f4beb33be539b09845ba5786
fc200e2cc35bd4c26735a9c0f04e972778df6e33
29012 F20101130_AABWPL fickett_f_Page_045.jpg
55a36d60c11bb9f10617ad0e0443216b
0f49f36d43cf16928c345c53c14aea9b3df5ab5f
2914 F20101130_AABWOX fickett_f_Page_132thm.jpg
c235996470930b6fb0de3b2e7150239d
509de4a72ccc960673c1cc835c827dde1ba164b1
23847 F20101130_AABXTC fickett_f_Page_036.QC.jpg
6a4348a51c73eb13de45ce487d17d955
616db24a396c6e708e5e8caf8378f400d568b9ae
6516 F20101130_AABXSO fickett_f_Page_019thm.jpg
6553e23bd1e3a33f69a4bf61f8d8213c
1b72f4423118a3ade055ef82f5038702983f21b8
25452 F20101130_AABWQA fickett_f_Page_131.jpg
66fac3b6ff141da87e3eed7804c3f987
e3e0a1fb6fd77693b9af13b164e9cbe41452c8e6
90164 F20101130_AABWPM fickett_f_Page_139.jpg
3d21c098f74c660a74b2a80a1eb3c108
08d77d48614f800aff0eb3ddf9c67475683e101e
6046 F20101130_AABWOY fickett_f_Page_026thm.jpg
9f473207c73980a11eb35d63524e3f75
0c1005fef1bfa7b20170959df08af7f0c336bcff
23178 F20101130_AABXTD fickett_f_Page_037.QC.jpg
7710b5ad48ed2529447715ac752dac5e
dafb72c72f81295265ce93cda499bebacc1781b8
6526 F20101130_AABXSP fickett_f_Page_020thm.jpg
4441c63a9b3421ac77ab615021944f9f
d0ed6fce76895ebefeeabf4b361fe117db5414fc
9822 F20101130_AABWQB fickett_f_Page_001.pro
09c28275dfe6ddcfc3b7c43086ec157e
7e709e7ed7aae59dd229879d8e9d1766c9b0422d
777 F20101130_AABWPN fickett_f_Page_119.txt
9dad15430521b2abd9b014470a20e42f
e0aa1b51a86e6a10efe11e5f6af9467a6341b73f
65121 F20101130_AABWOZ fickett_f_Page_132.jp2
ac38291ef0089aa9ca1f071af37137df
e17dc5f1a50aaa2957c43157e939f98392f2ef44
13996 F20101130_AABXTE fickett_f_Page_039.QC.jpg
012813ceb0a638dc9b57c7b4fe933b08
a4f0984d417cdccbe35329f36c2d02dc28139f98
23185 F20101130_AABXSQ fickett_f_Page_021.QC.jpg
d1ae1206390ec915035137e7dfac3fc3
9b576295563cc350f0fa5864b324a6d5b2b1d667
2520 F20101130_AABWQC fickett_f_Page_048.txt
f535a49fa4924508a4d268d43f3863e5
96e4e6b00508c50fdf9d36e01712f28f050460d2
726003 F20101130_AABWPO fickett_f_Page_113.jp2
83d74af5f637033fd2b3e78228bcd90a
b875ded736d5317f334a2f24a9c90fc3e9cd4daa
10409 F20101130_AABXTF fickett_f_Page_040.QC.jpg
a3beb2bf9b5a08b68c306ed2daff22bb
c33a9ade587b06f6bfe7dff270eb689b3a46463a
6524 F20101130_AABXSR fickett_f_Page_021thm.jpg
d62f0c4a8c100b507e1bec8090488ed2
2880a7cf9fdc7d594a35d38a7585926b9172f22d
92752 F20101130_AABWQD fickett_f_Page_140.jpg
fd1fa4a71a0cb6f7f15277ed3a1014fe
b55fdf26da9591b586a5f7302189b4f0354b947c
17538 F20101130_AABWPP fickett_f_Page_145.jp2
496935ccf7b2eec46f2f078ef16f404b
8cff73341af793656d65eed63ecbb9656cde88a1
3278 F20101130_AABXTG fickett_f_Page_040thm.jpg
246e8ebdfc7d0d9f4fe75a1ad7828476
2ea6e6a77399d861914cf94acbac9b9bd1ea5b89
24118 F20101130_AABXSS fickett_f_Page_023.QC.jpg
58b205e86f4312919d2a8178cbacebbd
d4102a14cc7155f703b7f241d7a598da2c17653a
1659 F20101130_AABWQE fickett_f_Page_012.txt
e26a1255c1cae8e35bc70f6644812df6
b9d7a5ae8b64449567f508c6198bc8fb04345e6b
110403 F20101130_AABWPQ fickett_f_Page_020.jp2
6bba15224a4b272e95d607bf73c08571
205ef85dde608163c4a29162b7d3cefd12119e2e
9182 F20101130_AABXTH fickett_f_Page_041.QC.jpg
8535d9b9c025c371d131c009e752a36d
5a37cc0f3deadf78559de2572cb981203b663858
23359 F20101130_AABXST fickett_f_Page_024.QC.jpg
61c82fd5ecd396ea1536ca0bf2f72dd5
d457ebdae4db231e6a1c81aadb2452af85464345
13946 F20101130_AABWQF fickett_f_Page_003.jpg
8e8333a084aed1622847ef9cec9ffb83
41a18d76acce8e345bfe848a360383f0e1a418ee
36324 F20101130_AABWPR fickett_f_Page_129.pro
3e820d898a7c05f2b1f010c1f6c2bd31
6fd3cf2492800f66c93bfcef28af384bacc41545
2869 F20101130_AABXTI fickett_f_Page_041thm.jpg
72629ea32026fb056bfd411a85a883a7
d494b685c64fa997d7c6d9fdc54fcbe7e388b20d
21468 F20101130_AABXSU fickett_f_Page_026.QC.jpg
c59967a353373f47d1b4dd6aa437b5da
fd956bb19710e91c6f43d6daf41be11c01091efb
F20101130_AABWQG fickett_f_Page_063.tif
e6e5aac1704c009ce431359d08b557e4
38a734896172e080b903a4f80592d974158a8d9a
71821 F20101130_AABWPS fickett_f_Page_072.jpg
b20f518e31fdac833659935e34838913
dcd16a66898af01855d842962f9f33236b34d391
9370 F20101130_AABXTJ fickett_f_Page_044.QC.jpg
219047ce840bbc8b03f194c2e40c9b19
7a340fdbee122e82cc56716ca8e83dbf71c2b232
6527 F20101130_AABXSV fickett_f_Page_027thm.jpg
ddec290c2614380d67685e60a0bf42a4
e55c4f0b0b0e77945880f8d8fd4c4bc99a4c4844
49112 F20101130_AABWQH fickett_f_Page_055.pro
541b2265dcd2610a312c521dfe70652b
b8e641b8b051b59a652ced11251b7a2d5b17e497
10669 F20101130_AABWPT fickett_f_Page_108.pro
a14655d86d9c22e0acdd7a7ad52030c7
1aad9a7018ac8e9ff765b22cd3c4963c4ebaa747
3089 F20101130_AABXTK fickett_f_Page_044thm.jpg
dc63cde99b0a7ca8e7bcfb7d4cc4b3aa
a27f507ee3c9e9b32addf2479c2c3b404ffc84c8
23379 F20101130_AABXSW fickett_f_Page_028.QC.jpg
d21f60e396f50a9897dcbe5e4a870995
988522f7bc49058be7392621ba17e9d933c686f7
23039 F20101130_AABWQI fickett_f_Page_033.QC.jpg
39e9ca6b116681dfb7784737d7496539
00eada08e755f752a0c704eb0bc1234093579ad5
2002 F20101130_AABWPU fickett_f_Page_028.txt
abaf0c84c231f5a6468b87aa280b14c3
f16ba710f1a04dd990edc180f2fea16e6985212f
9434 F20101130_AABXTL fickett_f_Page_045.QC.jpg
d7983dce8f5cc291a5779553e87e6593
9115c981518e629e34d645bbbc4634f5cf89dc43
6468 F20101130_AABXSX fickett_f_Page_028thm.jpg
dbc7f6eda81f3a771ba52687229b8050
ccf5436313d402bb2c2f3de9e3ff11da9515153f
23258 F20101130_AABWQJ fickett_f_Page_076.QC.jpg
30e9de2369836e3ca551f26c7a54f33b
d02f7a38d3fd91a864536f6e6393e6b372dcba0a
108481 F20101130_AABWPV fickett_f_Page_079.jp2
ed499d78a1c637eed1814c8a10cf2347
f60f8ecbd0fdb85d30762a8dc159f996cd83d4b2
7061 F20101130_AABXUA fickett_f_Page_057.QC.jpg
7efe16037e43743d74c9bffa48828276
65e20cfa834119e342b0b3257aa22728aef14d83
3015 F20101130_AABXTM fickett_f_Page_045thm.jpg
27b8c1cfb41b44765fd8a971b74111d0
0251d38c2a9b2e5c671a1ad31c86358a57fea4b0
6179 F20101130_AABXSY fickett_f_Page_029thm.jpg
d296e39838fe36abd0cb5421b10bcfa8
e101e89df66210bca62052c7eb188c656b7435b3
F20101130_AABWQK fickett_f_Page_139.tif
550406e0ff9aeb2d278f2be14e20ae48
c79575e30b2cf6ac588cec1514b04195b52e0044
110773 F20101130_AABWPW fickett_f_Page_069.jp2
cc511be401f95c7fb81f5bbaef372709
2f9ed2ed4f59bee45b481f1eaf525ae28f41eb3e
2345 F20101130_AABXUB fickett_f_Page_057thm.jpg
15df6150e2adb832d09bd77fc301caa2
59bd4cffdb4c6d7dd118fe480f802c819ebf2e29
10312 F20101130_AABXTN fickett_f_Page_046.QC.jpg
6d05cae1fe90b653f3c3eac802047a9b
b6acd49f97a86c602109fc1076e3e030ed3f0840
23578 F20101130_AABXSZ fickett_f_Page_032.QC.jpg
09d4eb030061b9e87f734b860edbf223
c111e5936d1d4baa2113ceb6aa4d4147db7af4c5
108110 F20101130_AABWQL fickett_f_Page_037.jp2
f1b36a034a19416f39a6528591dd12cf
05154f1fa4f281ecc292d7b3f8d636c9c4e85a9b
2942 F20101130_AABWPX fickett_f_Page_042.txt
8e139a201656309c520268376528c38a
7e9cea6a0b5fd84a859054ea0246cd7c43b4f19f
21007 F20101130_AABXUC fickett_f_Page_058.QC.jpg
0830cf2c12a108a5dcacf471a8eea472
d39e36daf12a6d49133a2eb5e22940ddf6a9026a
3166 F20101130_AABXTO fickett_f_Page_046thm.jpg
732580be68877c9effa7e842fcfa32ff
c779bec9e552f1cbd6c08910f919976571f3820b
10061 F20101130_AABWQM fickett_f_Page_043.QC.jpg
a842ad72f23f5d7214d6000bed546c28
27886ad547b5928e9267119c327bbf4c52fae5fa
49654 F20101130_AABWPY fickett_f_Page_037.pro
c9004497114ec2bc5dfd9641a09da3f7
4e0e2ee043e5520b79d70d751f91f6b3550c83bd
F20101130_AABWRA fickett_f_Page_141.tif
729055fe86d9ae971fc5ee767e4a4852
cfbebd63be1f253cc8091415d540652a2cc1a8b6
6082 F20101130_AABXUD fickett_f_Page_058thm.jpg
74a6395c340d22381b0ac5a675a576ee
cf28163e9513a298011bcf738ff7402154cf9cd5
9093 F20101130_AABXTP fickett_f_Page_048.QC.jpg
1926e4a229d63aea5ac40a5ce2533d3a
75552b851761058ea02aaa8fbc99cc159513ac58
6434 F20101130_AABWQN fickett_f_Page_079thm.jpg
b43e850992af4e993a958128e5d706a5
9f774d67f9a05a63a516c6ea21199ded3bc0b350
6392 F20101130_AABWPZ fickett_f_Page_080thm.jpg
a3b801b8325cd5154176cf040664ed4b
153e9e3ee1f3ff3de9a48117cf174bc51c7b9101
25271604 F20101130_AABWRB fickett_f_Page_103.tif
354ac203005a1774d3ad598be82377cf
0ddb3b31851cf1e26b16a802216ef921821a850d
23846 F20101130_AABXUE fickett_f_Page_059.QC.jpg
635f0a9b7548b108f6434ac1475e9ab8
a7d2ead832be8281c810eecf66c92cb46a596d17
9799 F20101130_AABXTQ fickett_f_Page_049.QC.jpg
094b99fbe9ea7752f4a2ba3f5c2ab5ca
08915663e59be1634d81de4b4a3815279d0eeb31
132235 F20101130_AABWQO fickett_f_Page_143.jp2
3656bbf0893fc75a1908d350eac10643
b702b0884f990520751d8d19034967d984c70f88
1054428 F20101130_AABWRC fickett_f_Page_096.tif
9c182417a48cb2436339e7f3d5a53f88
011405e5c6912be86e4c3f3f77e0f43e69e46723
24429 F20101130_AABXUF fickett_f_Page_060.QC.jpg
8edf0ce88c6452d4d57387f86f2ae502
ab628a2482a1bd1fddf491dd8d481cf1d963c1c4
3017 F20101130_AABXTR fickett_f_Page_049thm.jpg
ee9c697c4e0f5a5bafe724293243ab62
8c0e4eb2c2928ec06bee9efc1de84a62f7768cfd
7717 F20101130_AABWQP fickett_f_Page_133.QC.jpg
a1772ff483fd0ad1d32d9bf829aa7698
f51be4542467f5e383bc3fc2696a5ae461c357aa
1951 F20101130_AABWRD fickett_f_Page_087.txt
d7528f4240b0e4296bfaccfd07e6fb02
1e728af5c7f244413438501b2b655752361f7ce6
6737 F20101130_AABXUG fickett_f_Page_060thm.jpg
bbbdbb40304476c8765890a96af03817
085863a998ab787b95c099a9515ab35c3ca93c28
11132 F20101130_AABXTS fickett_f_Page_050.QC.jpg
df9ad83a47fd7d7cc970da757e4b07d6
080821295c1f373351e8579acf03f29c8accc9bc
2051 F20101130_AABWQQ fickett_f_Page_081.txt
e175c1263fcc6dea530cb1c9576f480a
b88c3119483d61f52e6030b3bed04e39652a5238
1628 F20101130_AABWRE fickett_f_Page_099.txt
f3c05eeef2e14c02dd3a7a5d36eb2f76
351e723876c0b612b3c83ffbd34ea8c82fe3615a
6569 F20101130_AABXUH fickett_f_Page_061thm.jpg
051717f80d2659d6d95fcd26f155cf92
f0ef8042f44316697cf7b71e368286f33883f060
9326 F20101130_AABXTT fickett_f_Page_051.QC.jpg
5aae004f43cf86c04be0cf1a1ec1ab62
1945eef7c877d17c0112ad63a6aa088aa9ba5b0e
50462 F20101130_AABWQR fickett_f_Page_013.pro
fd38df69a4337be090b57c59f9227706
c62b893c8f47f275619b9f7b5a712f25dda686af
1863 F20101130_AABWRF fickett_f_Page_078.txt
bb2caa97b35106cedfd520e5f985d80d
aea48f341701610581c3af74a95629f940f7bec3
23884 F20101130_AABXUI fickett_f_Page_063.QC.jpg
7f24c9308373bc3bed66379a65206fb4
6082e3d1d5964be6fe6a226d58dabf362b150ea7
2976 F20101130_AABXTU fickett_f_Page_051thm.jpg
18f652a34efab3970b6a569993470a3b
34dce46bc36fda6febd7630d8fcd6b149e698bd1
1548 F20101130_AABWQS fickett_f_Page_016.txt
7591fabadb055403679659dfe8c6b14f
7231d0c46735ec596994d05df8b9c8ffa6e4029d
F20101130_AABWRG fickett_f_Page_086.tif
526e419cad3e8d1560522433467b5831
6e881eb2939ca544c06c88709289be74a0d7c24e
6506 F20101130_AABXUJ fickett_f_Page_063thm.jpg
420e442c1619d38b053e6591e52efd6d
7cd6be952f53452c583b21ce9badf33c61a09e85
10053 F20101130_AABXTV fickett_f_Page_053.QC.jpg
57f424c132f1a85b943f78cdc4169f4c
a42c5726d32509889b5dd4b76393ed1d15024c34
15518 F20101130_AABWQT fickett_f_Page_008.QC.jpg
9cc707f69e3e7500fbed45989af3b4dc
40de513d6427588fe605d4bddac4f41e01e60a54
71865 F20101130_AABWRH fickett_f_Page_064.jpg
6fb10166f3d25935122b505e6529dae8
f4ef5ca7936c85410eea82f184e07ce10b6d8a10
22791 F20101130_AABXUK fickett_f_Page_066.QC.jpg
a20ff642cd055ef47dd92d6df03a4a2a
870ddb10b2fcd593b92dd0738ffc42dec4795f7e
3117 F20101130_AABXTW fickett_f_Page_053thm.jpg
ce184334202f3b3275b4b29e82e9c885
943e36559b4f7851aa03b684f2e1c550fc3ff98f
72054 F20101130_AABWQU fickett_f_Page_028.jpg
be204a381c2e6145caaac317127eca49
1df3e4180e56c079fecd35a1fddc915a22ce57b2
3568 F20101130_AABWRI fickett_f_Page_115thm.jpg
a2696811e2bfdc98f1cf86fa045ef1aa
a0b70e67743ec34613d18fc7256204a1d2b1d847
23154 F20101130_AABXUL fickett_f_Page_068.QC.jpg
e37f3d8d26f3cd424145c835d1def9b5
0abd5f853706680432c61844c6e7d403ad77f2fe
7956 F20101130_AABXTX fickett_f_Page_054.QC.jpg
4ca2e758e145753f9042838ee5fbaa3b
2039b55baa54c2bc21eda57cdcf7d55e8ea82607
732 F20101130_AABWQV fickett_f_Page_103.txt
3828eba9498c2729bc106b03c6870b41
f7d445ef256d418ac832eaedfcabc5280ee14d11
6765 F20101130_AABWRJ fickett_f_Page_073thm.jpg
01b81b0075c79eb76d9849cba95139cf
aa3fb5afcf25b160fce1e45540194f28853c207c
6522 F20101130_AABXVA fickett_f_Page_082thm.jpg
c35ee0a420574bade58ce0fd0f4babf9
8f54baf2ea8a3886894d87812a2eb869b5cbaede
6389 F20101130_AABXUM fickett_f_Page_068thm.jpg
b11d4e335855ebcf41a048ed8e9feeef
6f508f94d22aefa0d3d72647d7756daf2cd3cde9
2802 F20101130_AABXTY fickett_f_Page_054thm.jpg
af0a161975285fa1d8b7a047ebf82188
8cd4fdd54749c0e4d8e3313d705e61a518bd26b2
21901 F20101130_AABWQW fickett_f_Page_029.QC.jpg
d88f230de64f30df8fcba39e96bb5c0d
d6214ac6f05eb8dfb826065898f20420a30a859a
69346 F20101130_AABWRK fickett_f_Page_088.jpg
c3f0c390d3f2205ec469b2b4017ec3a8
7b9b8ab6bb610cfaf386e18fbec0b1663330f640
23326 F20101130_AABXVB fickett_f_Page_083.QC.jpg
036b3a26327ab6c1469fc73f2a589ce3
cc5954bf9862e91ff7384476988e334160dbadc8
24302 F20101130_AABXUN fickett_f_Page_069.QC.jpg
8bc38e3c43e591bda07ff5245d5ef36a
46941f386e02882b36e9dfc6efe6574628b9e7db
2843 F20101130_AABXTZ fickett_f_Page_055thm.jpg
7a60156998170d781482154f3333d6dd
4b06b3203dc079ba7387701916eb2c5eec2f31a5
F20101130_AABWQX fickett_f_Page_129.tif
a83644edbb97740708222694999fd95e
e4ce523db9fda0e356e5b67cb225623e56266e62
1879 F20101130_AABWRL fickett_f_Page_064.txt
b54085557df3d73ecba56f65bb137dc2
ffdb93e16353d08db48a055d4dfcce32afe0c998
23854 F20101130_AABXVC fickett_f_Page_085.QC.jpg
ef3b23371fffd907971e65bb76198675
ae94718f93bcb3bc10dd038bdf0bf7d7803e2376
24621 F20101130_AABXUO fickett_f_Page_070.QC.jpg
3fac84d58120aa8c1e439bab78fedaff
05b5539796c96fd55eb9ca4b2364c66c40c3bc0a
67763 F20101130_AABWQY fickett_f_Page_029.jpg
83c3866ed10a443538277609fcc57528
9ca165c5690835656bbf8640c5c7e146a98cfc77
78403 F20101130_AABWSA fickett_f_Page_100.jp2
8b06ead4930e92a1a42e4acd65b4ed6c
d0359f091857ffbdbe3b8043fa98e4f6e20535eb
38868 F20101130_AABWRM fickett_f_Page_016.pro
c2a26c032f3525276a8a2f5e07a7531a
28f5dd5a6e17217b283d2d06317a82ad506f6a6a
24284 F20101130_AABXVD fickett_f_Page_086.QC.jpg
9c1b9e98aeb84570a6e1bcb2fc013327
e5609c93c45c38871fe36e2122c78a84ebcd07af
6657 F20101130_AABXUP fickett_f_Page_070thm.jpg
5ffbb2c46a72dba9d0bb83059260f562
1d866d8ccb9cedeefc08b11dc16bf16761b61149
1618 F20101130_AABWQZ fickett_f_Page_131.txt
56625219b91bb2410a567f2b08b2c3e5
26b6b3bb7161c5a73b44212ace3aa862bccd4d36
1064 F20101130_AABWSB fickett_f_Page_039.txt
840986fbe5749582f0256c43fb99edd5
9008c247f00e3deb09ceba3b739ddcc578313754
6936 F20101130_AABWRN fickett_f_Page_056.QC.jpg
25936ce727877d946f745a7c55688e22
9d802bd3fccea07ea514e0c68c4466d0b5410f26
6664 F20101130_AABXVE fickett_f_Page_086thm.jpg
f71ef8da133c2f0d4769c2505ea91b69
1c7b485633b47bc23a7930136f0153b300614f59
23197 F20101130_AABXUQ fickett_f_Page_071.QC.jpg
6c21bbef0c7fa36f748d9c12bad8983d
9fb52940bc73588ab2cb06d529c21d4174d89d74
52254 F20101130_AABWSC fickett_f_Page_112.jpg
f89f501574bf034b083e697df4a7da49
c3496f98a194f2f711ccb60905249b24ec71b1d3
71929 F20101130_AABWRO fickett_f_Page_033.jpg
fc660a9e57af39c75b63c36d0fb0e7f0
21b66bfe0f408b148b48580bae9012bc02f772aa
6493 F20101130_AABXVF fickett_f_Page_087thm.jpg
149455c7e0f057bee33de5af53203d0b
78d106e12d2634eb519ab29a115d2318095a241c
23235 F20101130_AABXUR fickett_f_Page_072.QC.jpg
dd92ff0eb91690b1bc3f76d15425cc99
cf5e46fca5e0fd891d5d32db0e5ccdedb4e966e8
6880 F20101130_AABWSD fickett_f_Page_142thm.jpg
0688298e47f4341d6eceddb4a1399fdd
53eaeb154ea11e2030e1ca429d978bf020bd9690
70390 F20101130_AABWRP fickett_f_Page_078.jpg
b81021987c9ccbcb0b0f4d78bbf31a5e
a623194ed3af7e3e9983cd4af82fc85b66350a88
24615 F20101130_AABXVG fickett_f_Page_089.QC.jpg
e69eca56d7215436f79ec7bbcd17257f
c85db787ade74eb7fed61134c1b6c2e310b0f39d
24445 F20101130_AABXUS fickett_f_Page_073.QC.jpg
6ea30b80035b92d346c2f6ef210b03ec
77ea19b1f8502d8fd462f15f0fb95a3e38aa63c3
23400 F20101130_AABWSE fickett_f_Page_064.QC.jpg
ff1a2cd78ac7b5d0ff3c28e1007d8a1b
d0a9666ef32054f1c3a69d1680c1c8e77caaec28
47760 F20101130_AABWRQ fickett_f_Page_057.jp2
1a1d3f1920a5710250393f8d5c4dab51
0ebb59cb756947522b097038798f00cc159c57c7
24524 F20101130_AABXVH fickett_f_Page_090.QC.jpg
23f98b9ffaa1ce812416124aca2c6e2f
757e9bcbaf4d21335b2beca5adbfcb45dda53a7a
23573 F20101130_AABXUT fickett_f_Page_074.QC.jpg
7cec3d1fcbf6a7219d1c04b0b19590ba
b1db71e84fc6a2a94d822d9a7e686334376b4940
56207 F20101130_AABWSF fickett_f_Page_095.pro
e680dbf6108160cfdcf967340834a027
4d36230f489fde9d7b68788a6318210f7c9a2b84
1564 F20101130_AABWRR fickett_f_Page_008.txt
f481028b2bee1c5d29fe19bf171c9e27
c6f316035a0b8b94765d31a1e716085fc0e3801d
14943 F20101130_AABXVI fickett_f_Page_091.QC.jpg
af0b385b85ac90f242ba5fb7833a919b
2dc45ab5eb0f0999b9b8ff7facae5880b96eef44
6394 F20101130_AABXUU fickett_f_Page_074thm.jpg
9d2c03a5cb70a2cf4831a5c6d2ddb54f
7eda14b63a31a8969e8ecc02c09e1d03d5b4c27a
2987 F20101130_AABWSG fickett_f_Page_042thm.jpg
f44e5c35772a231403fc9fad4c5b1c5c
0e8ba6b8a0c9d5598d270173fdf11f3111a98f0a
3767 F20101130_AABWRS fickett_f_Page_010thm.jpg
d2fdab32177121bd3a44b86d3fbcab74
88b800ae82e97cf5307d12a9b2f9a17c3b055da4
4404 F20101130_AABXVJ fickett_f_Page_091thm.jpg
d9d6874b837b69a660c137b571f38ddb
356311741c2c541d70c14d6b4ca8e21aeded6b92
6490 F20101130_AABXUV fickett_f_Page_076thm.jpg
d796c77fcdf387922013c4d5b071901d
7ce61e0da6dd2abb9d827882e51c0d9ecdb93a72
108718 F20101130_AABWSH fickett_f_Page_021.jp2
9f0f7c8b86f264746e1693a15478945f
16844bc4f6236f2f6c27a6c772f86c62f9fd1234
6828 F20101130_AABWRT fickett_f_Page_128.jp2
835407f0e1cb8b69b0fdbd2d100dc31d
c0d5ff5441f67cbd8594b390dc9a412728d893b1
13332 F20101130_AABXVK fickett_f_Page_094.QC.jpg
c6a05ec04f0055729e2cbd819aee78c3
ca53ca5ca469c9d05140749041888a8f6aaf69f3
23881 F20101130_AABXUW fickett_f_Page_077.QC.jpg
6f0ad06d7316f75b34a437578afd1599
8e4590d9ba07eea768045434ba3f98b18603918b
87449 F20101130_AABWSI fickett_f_Page_135.jpg
fb5fb61234924837cd9e1ea4aa5bc8b6
9b2cb3dd6d6ca9d3c0c0ae3febfb742066e7c946
F20101130_AABWRU fickett_f_Page_110.tif
f17cc01f095f6b44414bf693047f85ca
094f1a2cc760bc9c5fa139c661d822fc9a8ec716
10494 F20101130_AABXWA fickett_f_Page_108.QC.jpg
972964d1a34b3025f6849e34236e3020
1f6af349c543c3293cfbecca7c6ab096d8a28061
4203 F20101130_AABXVL fickett_f_Page_095thm.jpg
f60f070926b87ec576bf257c70b707e8
924bac176cb32579792bd9e8332db9515ded9b10
6539 F20101130_AABXUX fickett_f_Page_077thm.jpg
428a8eb02e740217f86136bd8e1c18c6
032e51d7f8fff4853aee18f0c0212190abbdd3b8
F20101130_AABWSJ fickett_f_Page_022.tif
8f41dd76c18fa81fa6822ba293e5847d
2775588663afb25ca7e870e45b1114734cc34c33
6640 F20101130_AABWRV fickett_f_Page_035thm.jpg
abe48c656f1132f698c24ec01e230355
196d59d1cbc1ccae10c4d1367df743f2331eb3df
8665 F20101130_AABXVM fickett_f_Page_096.QC.jpg
51e23c0b4f18bf8540ef5e536cb4d7ec
bc1164035a1700a4de20a7c7cdbb29b3de52e69c
6668 F20101130_AABXUY fickett_f_Page_081thm.jpg
5a7ba47093407a63a2036085f44948fc
7cab7e70ef15547f0c4a1a6622bda3f62139ef42
60815 F20101130_AABWSK fickett_f_Page_011.jp2
39a8fd9130a71d62f823592356936a06
03d84e398049df32f3b61a0cb6d0ed90b191d638
11117 F20101130_AABWRW fickett_f_Page_128.jpg
79d72f11d6fb3857d217b1e472fa865c
9df51a91eeb87aaffbe590a389bc43f885a3c878
15859 F20101130_AABXWB fickett_f_Page_109.QC.jpg
6c5c9ee5fa104c762c7765522182e09c
49d141bade0a9ace611047d248ea8d5d1b5bd23c
2830 F20101130_AABXVN fickett_f_Page_096thm.jpg
76eea020d2bf331c5a74daa910020e10
1e7470ad1895ee23e562037bf011e2f08a22ceeb
23740 F20101130_AABXUZ fickett_f_Page_082.QC.jpg
a42d1806ccbf17a7623e6c6f9f7b460d
5411bffef6fa579474d80c0050717cd4fee308cd
77564 F20101130_AABWSL fickett_f_Page_042.jp2
a334b8a0b65494ed7be8acd6941f0519
b8a0e0d39b57c455c8b66b3f78bf0ab8a9fec26d
2032 F20101130_AABWRX fickett_f_Page_018.txt
c82a01bc180bec662b722c8c06cca994
2626510e1faf56bc1a1bbf6cd468939f89f33b70
15308 F20101130_AABXWC fickett_f_Page_110.QC.jpg
23db416e39d8658e6133dc8783e4af4f
29d186d93235abeaeeb296c70b942a32364eaf51
11797 F20101130_AABXVO fickett_f_Page_097.QC.jpg
be4d979cfd79a606df9b2b606a711ca7
09a39d3d6286c74b67b5ef00d9a2fc12b2bb7998
F20101130_AABWSM fickett_f_Page_119.tif
de75265ef38db949dc5bec893ad604cc
86e360b93251e9b117f9490f64745e18edd4bfdf
113114 F20101130_AABWRY fickett_f_Page_073.jp2
b28b8a558d5bd20b2f03738150ae82c9
fc28447ad15f78691e7c0b238aac24c929f4c91c
49690 F20101130_AABWTA fickett_f_Page_087.pro
5a793720ef1abe2973a23b2da787ee22
f6e6eaf043db8f308f678fad94ccd270cbd78df7
16245 F20101130_AABXWD fickett_f_Page_111.QC.jpg
e9587914aa57d3e96691b23733fef09f
e33998f9e8f27725c80bcffec5c8eb025773abbf
3629 F20101130_AABXVP fickett_f_Page_097thm.jpg
7fad7de41e68b8d810816ebd8d2b2c4f
43ff73cd552aea27a26aa6955a88d9d3a5d7c038
6409 F20101130_AABWSN fickett_f_Page_038thm.jpg
d938011e12b441d62ac88f2f872b7f1b
122153d6841384d9061462fa8c97a55edc075aad
6266 F20101130_AABWRZ fickett_f_Page_064thm.jpg
ceeaf8cfdc7e5419687201288014240a
37099c4e203c807e1fae65490405e46dcc1f0a4e
847 F20101130_AABWTB fickett_f_Page_105.txt
ee1b4c30a8714b26fa57a67bc45d9ba1
5dcca93db8735537c1479bae82ff5b9e37f55de9
4754 F20101130_AABXWE fickett_f_Page_111thm.jpg
1065b3a509b9b4af60dc458b88ac5554
be556635c217f92a6edc65e34cc1ebc950c9a837
9195 F20101130_AABXVQ fickett_f_Page_098.QC.jpg
1db9d6dfdb0b14f44f604f14c79839a5
ddc2d2cbcc8c9e8953753e482e01139b584d0c49
591 F20101130_AABWSO fickett_f_Page_108.txt
114f3e731303cc0d0e451aa3fcd15687
0d373d8617eb7d22e6a48cce3d2fa36ce74b13c7
F20101130_AABWTC fickett_f_Page_076.tif
72ea6d9ab9a1e6e2377da07b3ab34383
629a7e04edc1086564d6038bcb268a1343481f4d
15745 F20101130_AABXWF fickett_f_Page_112.QC.jpg
650286748e7788635e2e5a19315af8c7
a40028f8bc6cdf5651084bda0daef09549c45ea0
9532 F20101130_AABXVR fickett_f_Page_099.QC.jpg
318545d43e938e0790348f8bd499202c
f3dceef002ac297b236fb610a5a39eae5b045428
39309 F20101130_AABWSP fickett_f_Page_115.jpg
1c053a2f2d831bac4bcb1aad2769d8a6
ec626522fb219880c77bdcc763a33ace4ef476fb
32132 F20101130_AABWTD fickett_f_Page_046.jpg
c25c48d5ea1532e800ac6b0b37f47929
687a68a02e3199b3ff60608a09981a472aa02fd5
16694 F20101130_AABXWG fickett_f_Page_113.QC.jpg
89d8b7215d7aa7de8bb2180f3787f397
dc2a588f092ee2bfc190fb04af3f31e2b90e837d
F20101130_AABXVS fickett_f_Page_100thm.jpg
a3a8c76f628379174bf3b3565cdfdded
264e3c845b8da9c0a51cfe5151c7f9bea32c5891
F20101130_AABWSQ fickett_f_Page_113.tif
4557c495c1affa05aafe5d9ba5133881
24e7122c80e2a3f94720653cdaf7b9dcaf31d6da
28554 F20101130_AABWTE fickett_f_Page_044.jpg
9e384dfb3f41d82995e797bfde553b1b
c5a90b1dffd569cc3a50dae01e63a0cb89eb9f1c
4991 F20101130_AABXWH fickett_f_Page_113thm.jpg
7f2bf2f3905586373df9ec68738a7061
f2541d73c64b84ff271fedb665fd9abf694c7958
9846 F20101130_AABXVT fickett_f_Page_103.QC.jpg
f312a67b565a0e12543110bd81e29cc1
75d19367547eae291489ba4a10514b51d6eaa9a5
12569 F20101130_AABWSR fickett_f_Page_117.pro
775950715af01718861a152e3569d741
d6a41c32ebe6ca694f74d2f34c30adf46e327465
108944 F20101130_AABWTF fickett_f_Page_074.jp2
79e86f91e55774468073c88b7ac987d9
9429c4d5ca3ecdb1c007aaae01aa605d49eaed54
16956 F20101130_AABXWI fickett_f_Page_114.QC.jpg
bdf05cccde7681af3ec0476a76aa2e80
7a4db92534df6b97dd55f4c0c855ee732ff17946
3234 F20101130_AABXVU fickett_f_Page_103thm.jpg
fa8963312325969be8d528d79dd4b497
87c95a3141b39cbaf847dfcd794d5799ad3bab31
28677 F20101130_AABWSS fickett_f_Page_048.jpg
8a1e4c796fb3e608296cdeb7c472b157
ab5622945829ae1d9385d617322468cb3e1df8c9
11343 F20101130_AABWTG fickett_f_Page_107.QC.jpg
148a690940b56b7ea70009ee28a58da3
ac80bf06130a99371892dfba21c540774fc1fe65
4761 F20101130_AABXWJ fickett_f_Page_114thm.jpg
d27c422258a97cc8e7765f6824f1b109
76dfd5283960b63cc476c034bf82860a894c818b
3861 F20101130_AABXVV fickett_f_Page_104thm.jpg
d49bbc8d3323986a205aca70363249d6
a8f9ef50d997e9e1f246250a7ef85e047915308f
1581 F20101130_AABWST fickett_f_Page_133.txt
e5a37084fd2253132bed940fabcf2042
a5c978ceb80ffbcae4a0f105c83a2668ee5c185b
2605 F20101130_AABWTH fickett_f_Page_135.txt
c22767762251453c22dbb2a608dcc107
090f298078cb10c35d4e30e8d8fc821ba0aec1c0
17545 F20101130_AABXWK fickett_f_Page_116.QC.jpg
603bbc8c1702cef661823283b68f09a2
5368ac3c7d4d97322a03009d4c33db0e51488f16
12192 F20101130_AABXVW fickett_f_Page_105.QC.jpg
2297657e08f903af1231f2fb5c2cf563
3bbecdc73d17e158471bdad01b2f43c4286c4aa2
F20101130_AABWSU fickett_f_Page_004.tif
1ee1fdc6970aef7b1e554df7c7bcd8c9
2ee25c5d3d2d513478bbf9d0594ef569b3674951
1375 F20101130_AABWTI fickett_f_Page_110.txt
6f681a137316353164620fe33d9ac207
b3409447ee80cfd19a89b66b9ce5b488a6219318
8632 F20101130_AABXXA fickett_f_Page_129.QC.jpg
31741177e8d0fe1fb7b004a193f752d2
4fc743340db0eb1b392e1c63b4e9cacc9a5ccd00
5535 F20101130_AABXWL fickett_f_Page_116thm.jpg
c7c894a0bf2bfbb237822a0a6ac6f9a9
e74cf35c512394bf1f09e750779d17a515779710
3829 F20101130_AABXVX fickett_f_Page_105thm.jpg
130a692df4e0713d78583d4fe6ce8e3b
349b5702cec12c18b489f4d2e1777aa4af2bdbef
130148 F20101130_AABWSV fickett_f_Page_142.jp2
f55eaa8cfd5567fbe97af8f97eb8316d
53fd624f898c968d2c4dfb381d210017805b4864
611 F20101130_AABWTJ fickett_f_Page_118.txt
148eafd41bea1c38d480d963d6b15c9d
0138f9ef9c709b9edc6c8e45dda2b792649bfe3b
7686 F20101130_AABXXB fickett_f_Page_131.QC.jpg
87c0e2b6a95a6e9945eddf7d60b633f8
e5fcc2303823ceed01143f2a1882ef9e00131b23
11110 F20101130_AABXWM fickett_f_Page_117.QC.jpg
185236c99008ac94c27af139227dcf93
17fd09ed59e01a638f6c34fc81fc15068a9297f8
14213 F20101130_AABXVY fickett_f_Page_106.QC.jpg
06e767125f3a1594894c57bf26fac75b
c282852c743de303416c8962ea1a1a03f637a8bb
9147 F20101130_AABWSW fickett_f_Page_006.QC.jpg
58c3bf58b6d18029bac1a210df0bd786
eb2b23bae3ccd871396b2a5b3027e14c356efcf8
2057 F20101130_AABWTK fickett_f_Page_070.txt
21579bf69f17a01daccc3d83ec4066c5
d5386ee3f42f86951cc528c7a08dfd0642e86421
3486 F20101130_AABXWN fickett_f_Page_117thm.jpg
1ddab4caeee8ca4d6899e9ae17265da2
fc07230b89e4aa81b8a7da2f02bba04f4f1d352f
3718 F20101130_AABXVZ fickett_f_Page_107thm.jpg
eadce35ece65bf13a71fd916845768a1
bd3d1a4bf404eee2ca33a3f64c2b1c84a6e1db4e
110650 F20101130_AABWSX fickett_f_Page_013.jp2
1448a402f5b199ac9701df7bd8e2cfc4
3b9505c541d444dbe7d18b72cc0ca6ff305e01b0
2014 F20101130_AABWTL fickett_f_Page_073.txt
232073f2980ae9c81ddf626ec75a4424
ae9fba1f7d179ea9a3f4133967538f4db42b5316
2516 F20101130_AABXXC fickett_f_Page_131thm.jpg
58e2f905fd979062903df09dc51d32ba
3dec4f30ee78252a5ad7000ec0fde715f2a32a00
11247 F20101130_AABXWO fickett_f_Page_118.QC.jpg
477d5befdbcf4b94916d4e878441d01f
63a46f7128d51e2ef6874ce909d6576b580f31b6
22329 F20101130_AABWSY fickett_f_Page_111.pro
422b546cceea5174b5628a72008f7a04
285127c5e24fb2511689a38b52a43d1c5d8e0c2c
1149 F20101130_AABWUA fickett_f_Page_113.txt
c45a59373eef1e38ef0dd7d99099b7f2
4467dcacb27a5bd30a0e74bd4a0db4c522d1cb80
6519 F20101130_AABWTM fickett_f_Page_025thm.jpg
167f0742ca2a28a5e1e3674a6f3043c0
5ee05c3671f4831a377c8246ee708e96b61580a1
8921 F20101130_AABXXD fickett_f_Page_132.QC.jpg
29ff0893f19c062617a9b0975899534d
50bbf2570964cafaccf20663caac1a75d6c820d9
3519 F20101130_AABXWP fickett_f_Page_118thm.jpg
4e97c0c1ecdaf9a7aeb7bd822b2ccf74
14cb483dbd0ce406a0067c6784a8e98913700236
89684 F20101130_AABWSZ fickett_f_Page_007.jpg
736db07139db592ff0b446270375c4d9
291443b6b855efd15191c07ace3a42a5a7fe674f
24737 F20101130_AABWUB fickett_f_Page_116.pro
a24a167d59fe984c568cbf1b20bf0739
335bdfb12fbda3b6bd0e23fca358910cda84f8e9
110524 F20101130_AABWTN fickett_f_Page_033.jp2
2868104a0a8beeba3af4220b0a6159ab
830abafc9c446f62f68440d6dff4386a9242b02c
22087 F20101130_AABXXE fickett_f_Page_134.QC.jpg
fd6c5fe120b7df0663b5cda2e9dff8e2
c3efac347b6c10140b365fd76edb28366bfd6a87
3749 F20101130_AABXWQ fickett_f_Page_119thm.jpg
d2a67c9e1ea45e11b8ffacc1072cdaf6
5bf6b714366c2aec3494a8cc6a2e809417a2a098
F20101130_AABWUC fickett_f_Page_143.tif
bbe587e7dfbdf8f2d057689407ad606d
d30d8c175d0c6691ca9d88a334a3788d04880959
3141 F20101130_AABWTO fickett_f_Page_043thm.jpg
35b1594058699d8e801fa330c07da93f
b753888b9d578fcd92455414993045b1f5df3425
6293 F20101130_AABXXF fickett_f_Page_134thm.jpg
89e2a1c7101eb73c961e638c0c23b38f
363d831145ffe7a7f3a393454d44dd605a4a57b1
3329 F20101130_AABXWR fickett_f_Page_121thm.jpg
9a46fb8bb3cad37682ac44e5e951f6ee
4b8e40994934cc7b5118184b92c4c9a329bbf3c7
23052 F20101130_AABWUD fickett_f_Page_088.QC.jpg
7cf2c0364a69b89baa57f7c5a7c1ef2b
4da5ecfb25ebd06fa4d88001b36b69f897e3a282
64963 F20101130_AABWTP fickett_f_Page_098.jp2
4fc406c610f93a91829137358a0a61dc
cfbf218fda437c810f55c4590b8f3c9ea892ac4c
24165 F20101130_AABXXG fickett_f_Page_136.QC.jpg
532eda3cc15c271a19f127134857d062
f0b3f280c8733f42d87ca30a6c193ef2a825f391
9716 F20101130_AABXWS fickett_f_Page_122.QC.jpg
aed0724998bba35ec1e0db73adc7c5f9
cb78f83f75e381945026ee7c4e3da023611df8d9
35223 F20101130_AABWUE fickett_f_Page_117.jpg
17d44db6793ad4d78f7922b4b357fdef
06d4772b7ea8b2f6fa5d971c2baaacefed2a3b25
1410 F20101130_AABWTQ fickett_f_Page_011.txt
ca509a79870c16bb616a210f60ceab52
a5a31c11dc13cd96d96ffe0d17e10bcba4dfd5b7
25553 F20101130_AABXXH fickett_f_Page_137.QC.jpg
e77c55d607e040aff0b7a4f3b40d904f
ccdc203ea3ac2131914e1a7506c6295c4daa9382
9327 F20101130_AABXWT fickett_f_Page_123.QC.jpg
66e263143e4f4c7c62676cd25dfe5dc8
d349ebcc9fb9c3cd36e725bee8b0e52297e57861
36331 F20101130_AABWUF fickett_f_Page_100.jpg
bcec25a8272f7b72b8cd115d74f13c00
ccbac245ad895f4daf5fd5f5696870bd3d792c8f
32514 F20101130_AABWTR fickett_f_Page_133.pro
ccdc853b2b967bfe440f9d1921d9e459
0e1cfa2aa44b607710a09ad1ef5ccf72e894a915
F20101130_AABXAA fickett_f_Page_101.tif
d9b25a95ce50ee00c7cae4be5e4f8324
24c45ce68e5ef965ca1aa52f89058d3d0b13a4a7
26997 F20101130_AABXXI fickett_f_Page_138.QC.jpg
7a1f284b15951fe6c08e817f67bbafdc
460b2b8787ba052a6248e89aa2f5395d6c71b62d
3019 F20101130_AABXWU fickett_f_Page_123thm.jpg
4bcd107a6ca414e97480e8ba14f20eb9
2ebd665c86b030585e1e2406e51f481c7feac20a
36659 F20101130_AABWUG fickett_f_Page_097.jpg
db3d20e71e024478bb793dda81fb727f
66006e223039db7732088cbcb0177842c688c54e
F20101130_AABWTS fickett_f_Page_017.tif
b67a2b5a256c7fa562498fe8a0d84c70
543dcba3c6718938b6bc241a00b7e8d8b36e4acc
114152 F20101130_AABXAB fickett_f_Page_070.jp2
b8325f76159b77cdc69c44ebf42fe1a1
4b0ece2eb0ef7e779b05fa779762a31a9e7cb1bf
7129 F20101130_AABXXJ fickett_f_Page_138thm.jpg
84155938c71f6b83804c791b681c4685
6705781a57bd807c20b29f0d9025d1f39c1fb253
2905 F20101130_AABXWV fickett_f_Page_124thm.jpg
e39005425d261218e1e7f88b725f3ce5
89cce612b4e0fe2f434c0176c16cb29436a048b8
2061 F20101130_AABWUH fickett_f_Page_060.txt
a3f602caa057a171f046a05891942225
9afaf27137293734baf37273e4e620512a0590ff
74531 F20101130_AABWTT fickett_f_Page_075.jpg
6b00840b13e3e69c2cd8437b9afa9c14
0003608c58c04cc62bf3a1a0f00a9bf79d36cc94
F20101130_AABXAC fickett_f_Page_023.tif
61db8c0e5a414a01bdef02f896e5a6f9
27b1a05c6690ea75a88005ac9caf84f011764d58
25694 F20101130_AABXXK fickett_f_Page_139.QC.jpg
6954c10e7a04ce8cdb1a06e95fa264b8
672c66597cafba929d8e782f5df794b05ba42724
9492 F20101130_AABXWW fickett_f_Page_125.QC.jpg
79f51bb242a64a087b63df15cf794e53
68b43cab0aff679513e6440dd40e48b3ab0fedbf
10322 F20101130_AABWUI fickett_f_Page_002.jpg
b8938905256c2741499a596618345e51
aa9c3b236f5cbfbf3e059f742316383962da27eb
5949 F20101130_AABWTU fickett_f_Page_007thm.jpg
404c8e5036e71e488f0f63018e164d5b
a95b3e4264423ca7916e4d4e3b57d3c647e37530
2028 F20101130_AABXAD fickett_f_Page_086.txt
0b4048306548cd0b8188ad11d784ed11
5e57ffb62896d41c9092a0221c5b6178f96e4a02
26350 F20101130_AABXXL fickett_f_Page_140.QC.jpg
04a1d9968ed65e304cd14c617f08164f
829b9dd0f24a91efd4a7073e65350e1651ba693b
12816 F20101130_AABXWX fickett_f_Page_127.QC.jpg
ad05f6d08e355ba9951bbcbeb2c43165
34ca8e9e57eb6d500c87e916ba8d2381dc924bf8
F20101130_AABWUJ fickett_f_Page_016.tif
d718861ddb4ef01b227683b182c7aa14
5232bf1cad6dac08732b60f2e7a0be39a3bb5df5
327 F20101130_AABWTV fickett_f_Page_145.txt
f12fdcf15a74e5b8f64fb127e9ec95b1
9175ea8e540446181630ef5e81274314bfbdba7c
396437 F20101130_AABXAE fickett_f_Page_101.jp2
55dbf635b5917b68619375ac3a837de5
15736c84b2ec694a3fef9e83769cb9009e2df5a3
6995 F20101130_AABXXM fickett_f_Page_140thm.jpg
a535942b584ff64f5846d23f03fae1d2
4bb0a2c215eb85ac9d09eead01189ff57d4db1e1
3603 F20101130_AABXWY fickett_f_Page_128.QC.jpg
62d66fc7f77acf3fd7b2a20f67a365d4
473691d5312bb9b0ae2d18ffd2913991b10a0b86
73750 F20101130_AABWUK fickett_f_Page_091.jp2
6ddc68038dd66f14003108db6f65d969
ba314e3a8bf0a2eade4c4af8e4b88e4bab5aac6d
47304 F20101130_AABWTW fickett_f_Page_067.pro
d917e26406874dac411dc79241603dd9
938e4f5a94e6676a7983f4084ca31c67640f02a4
20792 F20101130_AABXAF fickett_f_Page_057.jpg
1f7174ce93f97bc76aaded0a889e372b
3780f3367ce9a7bab00c77cf04fa87966069d5f9
26168 F20101130_AABXXN fickett_f_Page_142.QC.jpg
9c23cc67073180044fc300f8c40a327d
4a9998456b7e40ecceacdbaa49eefcb58e36e210
1395 F20101130_AABXWZ fickett_f_Page_128thm.jpg
986e76972df480bd24acaa7bbf9d183c
6f610751888250674118d76413035de7b4638733
72664 F20101130_AABWVA fickett_f_Page_018.jpg
af12ccc36fc3b5917eae9f9fb2192bef
f4154b70dd2bc76e64963f8373dce6af395bb003
24573 F20101130_AABWUL fickett_f_Page_031.QC.jpg
01d4818c38dc01a9ebf8dc50a7a280c1
aeaca25845ef9a1010de840c9055404c7fc846de
39445 F20101130_AABWTX fickett_f_Page_132.pro
9084f3b2a351d1f7aec7bb558074ac6d
f8edac9de7388ae5a8c2b8cc62b22de4fe74391f
F20101130_AABXAG fickett_f_Page_029.tif
71cd9fe69b36ab7f5e41c453baa97dac
c0a731dc33432886a1f4593277d02df27f6bdc69
25526 F20101130_AABXXO fickett_f_Page_143.QC.jpg
af852d3be081803f37d89971b9514a26
c9fa21d3b5af95ab083e8e36bd5425107bf693dc
110072 F20101130_AABWUM fickett_f_Page_063.jp2
d0c9181a1a94864fbac7f85b5748797b
643ba153457ed24a2c2228e81278ab0a5d6ce481
1950 F20101130_AABWTY fickett_f_Page_021.txt
d24f8d3b68a2879a21dc5e0b78ae4e67
d57c2af108a7da7cb51dbabce1d55ab4d4581f82
50649 F20101130_AABXAH fickett_f_Page_036.pro
8bb83ca96daa7adc4b619474d78da866
00c16d3d87fbc554655a225c20802d31d144ecf4
25893 F20101130_AABXXP fickett_f_Page_144.QC.jpg
8ba2bea2178b1eabde0eef7e7459013e
5cbf32fb37f0d8b2da82119fdfbd96f70ef6b0ea
5328 F20101130_AABWVB fickett_f_Page_145.QC.jpg
dea5f7ddfa2776d3a9e839ba319121eb
dd58fa7cf87a7f9db88cfd523396b59ebc376b57
365567 F20101130_AABWUN fickett_f_Page_103.jp2
558efb7f09ae932f280962e586a83439
9bfe233ee9a754159d5eaa498875c16efd7a71fe
57811 F20101130_AABWTZ fickett_f_Page_046.pro
b3e6a2c1229fe6a66d94eb6ff21ad37f
49888512f22f1d698be32e9b825f1e56c4d13e02
2242 F20101130_AABXAI fickett_f_Page_097.txt
265d18b369a7da04df932a52ad1c3446
1f690934092f0163969f277c2e95c9bf78795828
6972 F20101130_AABXXQ fickett_f_Page_144thm.jpg
dd7bb89ca9a2802f56b0495eb2417023
58b54c51bb15f259d4d1d5f03c0736eda5aa2040
2480 F20101130_AABWVC fickett_f_Page_130thm.jpg
7e02f625d72a6ffc8f6501b8d64a6bf6
20545b9bfa2f6be0edc8dd7c979c8f0af3ab19d8
F20101130_AABWUO fickett_f_Page_089.tif
d0b5310a9385505c5d8b274b26da3dec
71a89a102c3b84376338da2e2ff9aaedb4a993aa
2033 F20101130_AABXAJ fickett_f_Page_100.txt
3b2b3556aea60f1e3b2cde014ea4a7c0
bc6cb0c3072ec945cd2e235f3ed49c0216b161b6
2666 F20101130_AABXXR fickett_f_Page_146thm.jpg
3b1dad10c1f13e0ce8a573a2442b5f94
1559d07f0ac472e47db31062ad086671b12087eb
1939 F20101130_AABWVD fickett_f_Page_065.txt
217b884a667de039ced24845a99bf9b4
debcfcdd19a542df7f1c3ff51e922f225c5d3c72
1919 F20101130_AABWUP fickett_f_Page_079.txt
0cfb4af2c81b0e938b401bfabf958448
2dc9172863f68a06c2181741a63cb0adb76c4626
3008 F20101130_AABXAK fickett_f_Page_048thm.jpg
2ab5478e71eaa8cbe99b8c6eb6fa002b
8fbe0ede32b0ea1a0de818eec98c63031e94617d
169298 F20101130_AABXXS UFE0010490_00001.mets FULL
896306dcb333e60577d3dcca125d46e8
1de061b513de730c54857a5ea792601430035a4b
3189 F20101130_AABWVE fickett_f_Page_052.txt
53b15ae560326fd34c4c65a9c97ac1e8
5f2e7069952f38f374759b8a2efa6bdb0088c9d6
1167 F20101130_AABWUQ fickett_f_Page_106.txt
5bdcb1f3438d8c087a6d259a992705aa
fc61993f85ff5e541742a909f6c8b128c41d2927
28852 F20101130_AABWVF fickett_f_Page_041.jpg
e23d872994ca12069a7a0b7f3271a085
f5b9484b0b2fba5a0e36dbf8affd7e74da2b464f
F20101130_AABWUR fickett_f_Page_105.tif
02d453a507dd50db1a18d1cdafd56b97
df07f445d6a0d83a4a38cd73f0f809e7010b8180
28774 F20101130_AABXBA fickett_f_Page_132.jpg
78ba1db1b1b148dcfd7ac91728e5878e
55a535a6bcc0b5f4f0d03f0a64a3158c01f1226c
F20101130_AABXAL fickett_f_Page_055.tif
37cfd1765707fb71093b25e5401cee32
b175b3a1c5f2c7e67ff3646c9af1b6add4fefce7
81518 F20101130_AABWVG fickett_f_Page_052.jp2
c42ab41fff121a5ddfb2fe7fb338295c
d7f2e5e411b7218f202da6e8096b6655b7b257ab
39257 F20101130_AABWUS fickett_f_Page_092.pro
47e9f8ac7420da384f05b63d8b40021e
c2f3f7c3cd0834d3980bda450b321179aaf4995f
22630 F20101130_AABXBB fickett_f_Page_067.QC.jpg
4b3df0a23cdb1e4de3eac76e1b933836
dee917937bcd2d6f634dde3b4adac9a7e41f4295
1014 F20101130_AABXAM fickett_f_Page_111.txt
49f04674b4aa003d311777d0bd51802f
4cd2888775ffbc07a69936acc6b1327fca7ac79d
1051982 F20101130_AABWVH fickett_f_Page_008.jp2
c790b7b33f2660573e6712a054d11434
9ae3402bb429dc81cfbfc49fd45db0a7cd698730
104226 F20101130_AABWUT fickett_f_Page_022.jp2
e8ab73eb8c76f909883e7b639fad4f79
cbb91a8e092a14794566fae62df00d4c90e7f431
35479 F20101130_AABXBC fickett_f_Page_118.jpg
7785cbc1f088eec45473d05a1cd275a8
e70bb1edc2ba7e7d528926384972b78aeed18f38
22893 F20101130_AABXAN fickett_f_Page_038.QC.jpg
d382e18fc2f4367d3cb07492d14a4753
96e269dc389b66f4d8e532926717572bb98bdfcb
F20101130_AABWVI fickett_f_Page_098.tif
13d74374ff9398926a257663fc68a89c
08039b30db5c948768a08082ca96ddd65bf755cc
450325 F20101130_AABWUU fickett_f_Page_115.jp2
19e50e75ece156c6fdcdd7adedbaa0b1
94fe4b3deedc79d677a031a3a0bd31dcf8ebe572
75605 F20101130_AABXBD fickett_f_Page_073.jpg
2023d96cc816643adb12e33fd947a119
f226c06afd157ff655f50845a82ada4fdf03b30b
2971 F20101130_AABXAO fickett_f_Page_040.txt
8bb1b65e0e42a91533287d099b0b72da
4ae85cd2d7bb229ecdb37859c1a0272352d622ae
6725 F20101130_AABWVJ fickett_f_Page_090thm.jpg
235bd28118d0f66de46af7056b743818
5b46f96a1cec1d27fc171bd1af76f868d15f5d1f
50135 F20101130_AABWUV fickett_f_Page_047.pro
621467d98c453d50f9c60e1aadb95516
c0e78cdedde021b20780303761f54eb1a2fa195f
131683 F20101130_AABXBE fickett_f_Page_144.jp2
07359ded54b32b33162bbe78d222f1bb
8f622af72e9ba8aa327ed1f892ec8e2b8bf5fb10
6727 F20101130_AABXAP fickett_f_Page_059thm.jpg
612188670eb473c8632c8daff9321ab4
9eda9a484d9e88b507f6dad29414fff189662e33
14878 F20101130_AABWVK fickett_f_Page_115.pro
0d59418d2ac42740b505314925ca1f3e
7127b5dd75b5c66d55fd6a1d5b66babd931c6af3
3912 F20101130_AABWUW fickett_f_Page_106thm.jpg
6cdd9e2ce0a0ac0152bf4a1691a15287
23300f8f2a717c3ca8df47bfb7c570e464093e47
8849 F20101130_AABXBF fickett_f_Page_146.QC.jpg
cd5c98787fc5c7062168bbea3e578398
82048e9b95d72a7e80f1331332746cc94b131ee5
11642 F20101130_AABXAQ fickett_f_Page_119.QC.jpg
dc9ae2b55ed71b043a6453c0faff9d50
3912a4dec38edf709ef6ace5f39d1950d66707fb
73364 F20101130_AABWVL fickett_f_Page_082.jpg
874b3e284584d321c52dc90de7a2c4cb
216dd1cf6aa8c855a0004e199f48fbce1e9611a8
41387 F20101130_AABWUX fickett_f_Page_104.jpg
f8496028d7b2efebdda8d10e374aea52
95bda1e54402020877e059f9edeb125ca89be6b3
43117 F20101130_AABXBG fickett_f_Page_100.pro
7eaf0cff6d5796057fdc6233fdc69089
9b631c39600e086f0ee9e5be4d344d4760566771
24233 F20101130_AABWWA fickett_f_Page_030.QC.jpg
6fa966f7a0aa5a84ff52b136b804feef
20eed97192257298e4b80c15634f9231c31d9c2e
59682 F20101130_AABXAR fickett_f_Page_012.jpg
075b7a1338e143a4eb206983de0f6007
7ebf3ed17bc0c811d03159b8db4cb08c45b838e6
F20101130_AABWVM fickett_f_Page_059.tif
8f482a797c7fab2e80023e6e7cb93606
475f4f51b45ee5d189b56f35a343be9641a3c7c6
585 F20101130_AABWUY fickett_f_Page_146.txt
3df54ec9a461dcccfb7f77a3d1e5ce32
021719538dbc496da543d4c99eec0a6a3ba03a87
11160 F20101130_AABXBH fickett_f_Page_003.jp2
4562ac808473ecd32532e8e4f102e0c0
5e5b40fa4cfa7ccc3ae4417a17bc9b80cd75a01f
4721 F20101130_AABWWB fickett_f_Page_110thm.jpg
a25ef67dcde5e652378f1beaa288d0f8
143d50e2d186e16f1a5969be96bf18c03f4e8641
6466 F20101130_AABXAS fickett_f_Page_024thm.jpg
bed5a6330aeec1ae791ae78a44d3a7b6
a7562c8f3f8e429a2bfe1f6ea25795fa6e129faf
3870 F20101130_AABWVN fickett_f_Page_094thm.jpg
111bb87cb2045f4c61592a485ba0a0b1
b7e36f926e305113022196aa283432a89a2226c2
9412 F20101130_AABWUZ fickett_f_Page_042.QC.jpg
9cfc262bcb1b8001c822bcc37853b8a2
018267d995d3f22d36f31ba824787d905398e0c7
F20101130_AABXBI fickett_f_Page_039.tif
67ce2b20347756d63cca223a9e685286
5e11db8229c8685c87d04d501b35b1ed3a623f51
10277 F20101130_AABXAT fickett_f_Page_101.pro
7e047180d19cd77babd418691a84a015
47089f2972cf484f52a6633c3594ac29395e8d96
7052 F20101130_AABWVO fickett_f_Page_141thm.jpg
8ec98508c66b48194a9c34d6bb17c411
a994266c1411d5a6796e97035626107815ee203b
F20101130_AABXBJ fickett_f_Page_125thm.jpg
5940af47d687bae2089c95e5ba6826dc
0da8f767f0695f22481c514b22dad2be49e1a8ff
5063 F20101130_AABWWC fickett_f_Page_120thm.jpg
53155f2f83cd0e92735b6e33271ea35e
22ab058c26dd570f857920a5ce8509f2b2c559d3
4159 F20101130_AABXAU fickett_f_Page_092thm.jpg
343ec78640b56c748ad5b1868c66dd9e
33d479da75e184c460f2f6a9753efb49f6a34850
6661 F20101130_AABWVP fickett_f_Page_030thm.jpg
eaaedef499ea3ecabcc15b2a68cc0b48
588ca7d34e6e402273b3b99213748e3b991923a9
63584 F20101130_AABXBK fickett_f_Page_007.pro
9cf1da1a37e1a4ffa857299d9fbc2127
545666f0173a343b62034991cc7f73640edecccf
45764 F20101130_AABWWD fickett_f_Page_029.pro
bbcb6e848afbf5fc1414ff91a8a5feb5
7b4aad9300f7847cc473ce781756ca0fcd2d1fb7
49619 F20101130_AABXAV fickett_f_Page_021.pro
225907ee95bb28649bf729002625106a
f4c6b82777cf6f3882d484de82f560121cc03405
6706 F20101130_AABWVQ fickett_f_Page_034thm.jpg
523d4ba13faeb25521618ee38880f632
0d016f166c530c0cdca7c85795067fe73c3d839f
6615 F20101130_AABXBL fickett_f_Page_072thm.jpg
b0977b7b8837e6d4a202998ac4ff39d0
cd13411bf34133382adcf05394111ddb6b9d454b
57156 F20101130_AABWWE fickett_f_Page_045.pro
8396e209e36b3d715f8b672762288d21
34cffedd8774d516789b894d837bb13ca1e6c132
F20101130_AABXAW fickett_f_Page_088.tif
5842b97b9316a9527a7287fa825407bb
567640e78d421fc0e400a149fe71356e153791e1
50593 F20101130_AABWVR fickett_f_Page_019.pro
f87d36f1d713440719b4722e8cf205ad
e201dcb1e357d244b320855458205d8f1a0babce
50058 F20101130_AABXCA fickett_f_Page_075.pro
be0d74648ba2f19b4b68eb99ac35f064
5a4ac407409cf8e2e32a72dd6f54ff5b2160b58e
F20101130_AABWWF fickett_f_Page_036.tif
a93a1d07ef77873c1d75a641ee03c0f4
82933046f5b0805e8040176b065ae145d0ae5cca
3128 F20101130_AABXAX fickett_f_Page_101thm.jpg
44909b1e3c6000012b714d4f5d1f7906
bd5bc757607b892f398a40acf2c3b478305f135b
4647 F20101130_AABWVS fickett_f_Page_109thm.jpg
bd44a209709b6ec9003b96b95b770961
b27858d0a66d42bd64dc2324dae36d5431d1c440
86 F20101130_AABXCB fickett_f_Page_128.txt
b3bd2c0839e3293b8c27cb9b6c8558ac
b55dc7f8ce555b12b3def97c9181baebcac9265c
43962 F20101130_AABXBM fickett_f_Page_095.jpg
25f42a39491fe647f805aba79fa302e5
04e80b4e0989f991499e5cedfdfefa76a0461a8a
2696 F20101130_AABWWG fickett_f_Page_138.txt
c5ad93d903aad1cbcd62568cdb5774b1
3223c192efa14f72bdb4a5a01a4cc430b5a6d24b
2011 F20101130_AABXAY fickett_f_Page_069.txt
87a2125dfb55946af6de45f2b50d8010
2bf936179a68f1c7ad13680c1da3534fc9fbde27
111583 F20101130_AABWVT fickett_f_Page_089.jp2
49f72246bb1d64d44fd4dced764d7815
403e23818ae552281ba2ccb163bec1574182a662
6376 F20101130_AABXCC fickett_f_Page_022thm.jpg
978e0855b0c67fb608ea710d61001bdb
b243c3074a42db5fbbf630dd6851f27ce26f0876
6832 F20101130_AABXBN fickett_f_Page_137thm.jpg
be8eb3d48cb2c939a712b7c62a1475fe
be002f3936ea75de68c952ec23e140144db0023b
34661 F20101130_AABWWH fickett_f_Page_126.jpg
f9862bd9f76044f5cd2498a81f865d74
1e6399d7c3a6be72d9d2e85db5c9e5039a8994a4
72721 F20101130_AABXAZ fickett_f_Page_087.jpg
717934ccd4243acca97d55aab483248a
01692311c053fb86e3dd1cd0604ae1970d6d745b
F20101130_AABWVU fickett_f_Page_102.tif
aceabcaa992aed37a660dacd3ae004cb
47e83345b483b287a2b840b588de6198a092899c
219459 F20101130_AABXCD UFE0010490_00001.xml
ed88d7d4ae45750afff5b591f23e17a9
9b56ea32dce1ac4c13511e91f75515c399f32f08
12843 F20101130_AABXBO fickett_f_Page_093.QC.jpg
cd01c825f82f762dd99c0eb236e635f8
0e080b66bc4eba77fa3bff912207ff504cdb97a9
6605 F20101130_AABWWI fickett_f_Page_075thm.jpg
b4faacd2eb47e97f7c1e1830e98225cb
cadc5be47e49f3804547010cab6c2e6cccc25c85
74542 F20101130_AABWVV fickett_f_Page_060.jpg
ad5c04c2285b92f4d08a3d028abdd25c
5165bdf14bf4dba66fc567f19fe57794b2e7935f
6689 F20101130_AABXBP fickett_f_Page_083thm.jpg
51900b80b3347c8ae34b8e757096056c
be0ebb83ab71f9b0fe69faa228659d8bfd347cc9
17847 F20101130_AABWWJ fickett_f_Page_120.QC.jpg
eb550490883572173f6180e53bf1f360
0470715a43a96451195392e934fec1cad15ea8f1
17770 F20101130_AABWVW fickett_f_Page_127.pro
7910ed27db010be251cb491daefcc559
f66fe1ce0822a54c8ee78093c8c85780f42bfb93
50588 F20101130_AABXBQ fickett_f_Page_074.pro
4689c121e598fa594035a8bf40d4df6e
c669fbd57b4ce3d806dab8e5692b11a9e67bee15
F20101130_AABWWK fickett_f_Page_077.tif
0096c12c5cde1cf78be195e32a200942
eed40f0b3da265addfac36d2e47ee3887cc1c134
26303 F20101130_AABXCG fickett_f_Page_001.jpg
077835a888965546d5ac627ce98bfb45
0d15d87cdacc36e5ebcb3e6eebd1e80b007b9716
118885 F20101130_AABWXA fickett_f_Page_136.jp2
ba0ce9db1071aa6e7bc1b9ca4da1a01b
0ec935d0e42c849b75c0fe40aa700dc85be2acc1
86198 F20101130_AABXBR fickett_f_Page_049.jp2
bb4ac551d6b528c31e197f6e4184a354
787cb690a7a2eaffac17e257875ea3b08b869766
74895 F20101130_AABWWL fickett_f_Page_031.jpg
ac911bf145b1481b889fb01a8f8939ab
aeb25b311e28fe03420d01c65b23b1dbc4c574f3
109904 F20101130_AABWVX fickett_f_Page_134.jp2
8cc736d1ad7290dd2e993b0414364655
d9b09c971d1ebfbb1164c7aaa694ae57130232da
70728 F20101130_AABXCH fickett_f_Page_005.jpg
070d017edef6a8c9a36b28d45022d5f9
312460f510fe1dee423910fea711b11be44100f5
4033 F20101130_AABWXB fickett_f_Page_003.QC.jpg
2cf9d517cb55a19a451c2ce19da46a99
ad20fe3ca2462fab5dd0ba4b7e9f6cc65c98b8c9
70317 F20101130_AABXBS fickett_f_Page_071.jpg
5552ac7b337100f10bf57ab3cc19aba5
9bdf3b890cbcf63686cb4e6787c03bda088f63f6
26864 F20101130_AABWWM fickett_f_Page_055.jpg
23ec45679627a1da8b18e8fcd3824389
26030a7dd0208f166738d45c2d4f6dde1b431e8e
F20101130_AABWVY fickett_f_Page_037.tif
2c90945ad1ea6d942191db4a306dfb28
0e0aca92850f30ab7e4bd4210d08632117866628
34748 F20101130_AABXCI fickett_f_Page_006.jpg
26a5ba92ba0c7eed26afd583b2e558f5
3a3df5fab9964485a6e42f881103569ca7b8a707
11744 F20101130_AABWXC fickett_f_Page_115.QC.jpg
2ac9af25cabc746ba8804a027c194fe3
899abb9400af0810228e080c25749f9cbc8b8b24
109712 F20101130_AABXBT fickett_f_Page_028.jp2
ef8ecb98ea76fdebff9b036d4e20e983
e073f8e6c1092d812992fae7ee25a7f160871a50
649 F20101130_AABWWN fickett_f_Page_122.txt
f8241aba68fc891ffa449bc5ad7582fc
3ed23160ccced152e2344049f4a409a2dcd18cfb
22390 F20101130_AABWVZ fickett_f_Page_004.QC.jpg
9ecfb5c37146a7253710ac662b00c7fd
cd865a707d54214ea05513d935a9bc43ca2f626c
57147 F20101130_AABXCJ fickett_f_Page_008.jpg
99910a5a85f66a51b4b20ca81d73c39f
29a51f425e35b68cc004b462b5b3b491bb7790b9
F20101130_AABXBU fickett_f_Page_009.tif
068ebe8f9dfc9be6dea612d541c736aa
d81da9c4252ec05daf06190e7c0a31bbf0ff4c24
F20101130_AABWWO fickett_f_Page_015.tif
55c8181f8196785c524e7b5f61561bd3
73e1e53863343acb34fc0f02397543a2ab7bf889
78333 F20101130_AABXCK fickett_f_Page_009.jpg
354f420de936ff12dead536159b309d8
7da26f434e25955b9ff67c7c8bbbbc4346121981
6528 F20101130_AABWXD fickett_f_Page_069thm.jpg
d655528294f570e01788da28904c8d44
c0e29b6ccd2c0b48a6474dff679d4cfd3453522b
70605 F20101130_AABXBV fickett_f_Page_015.jpg
492a9b945ab80649c945df132bba1696
6e6b47a8d8d12de0423a17595383bb1f32fe6b6a
92034 F20101130_AABWWP fickett_f_Page_094.jp2
599590c95d7d730779941e4c4f409ddc
382c6662a205d13034aa49ca73698a01436e8133
43734 F20101130_AABXCL fickett_f_Page_010.jpg
a9fc6c7660c80ebaeee685524a7b6eec
7ad5cc725a83e2ff2896c1df4196cd7cb1ace2ae
26220 F20101130_AABWXE fickett_f_Page_141.QC.jpg
618aacd7b83ab6becab1ec2791c06c64
e4e7405bf65b8085afb33c827438535e9af8c780
72364 F20101130_AABXBW fickett_f_Page_020.jpg
81795eef287c41292ca93aaadad264fb
784926fbae1c9a8a9fd753ab06f703fac1c3a05b
12905 F20101130_AABWWQ fickett_f_Page_104.QC.jpg
a12dd8362884e8c0217f5e3316ec4cf7
02aa029ef9eb99012b985710c91293976f48f24c
42864 F20101130_AABXCM fickett_f_Page_011.jpg
b43e2a6eba336d1a13712d973aed6d54
27d7edb4d51e989317e5c65209260ddc62ac4ede
3324 F20101130_AABWXF fickett_f_Page_043.txt
207ef4e6d7742046c25ade5340db884d
7df28ab36fd083c3d79aef8bc720865f66917c81
83135 F20101130_AABXBX fickett_f_Page_051.jp2
c89fd7dec6519521f2da471b587e3100
23b4b1070ce92fc7cee0c8a550edca4074965398
6420 F20101130_AABWWR fickett_f_Page_065thm.jpg
f1540cb25ed62fdd95107358da31a664
e1f5f0c6845c24fb1af9696e79c20ca24a480ee1
71147 F20101130_AABXDA fickett_f_Page_037.jpg
abc672a6ce55bf26d2f14a0d6cfb30f9
24ea24d39dce22b2084500ff5c39cf272a25f06c
22151 F20101130_AABWXG fickett_f_Page_009.QC.jpg
540f39a8b937256ffc4fa1a743c7571a
114c4e8b653577ec0047e0292d1f8fca84da2016
6416 F20101130_AABXBY fickett_f_Page_032thm.jpg
5d3d5bcd901b683be2a016f95f35ec5b
a59a4bd0b07e02b23d5dad213576651c943adca1
522494 F20101130_AABWWS fickett_f_Page_105.jp2
9d7f6c450dee745f43cb98f95d6ac4b3
ce59c9fa5f3090c8fa7d56c524ce0527668d9596
69938 F20101130_AABXDB fickett_f_Page_038.jpg
990af0d4e1993d469729e6000a83b1de
a2d09d42e0969c197004c7b708a22d339ec74a6f
73833 F20101130_AABXCN fickett_f_Page_013.jpg
c9cae8174a6b932fe62375cfc5b45311
9b1eadb59f574e27dc322a264dee73bcb39141a0
2718 F20101130_AABWXH fickett_f_Page_055.txt
c5abdf4d224562a036c790b4aff3aed1
a81c03db49909877777fcdf704ed91174007fe0c
23602 F20101130_AABXBZ fickett_f_Page_079.QC.jpg
abbabf9522be7c70294a624f21206c83
b5bd749b84be3f613441982a94bf4ee82e12bcf0
24244 F20101130_AABWWT fickett_f_Page_081.QC.jpg
7ba06141c6fa503f4d2199901296e191
c295deef5005735f3a8c31f09cc445b01d51ecbb
34804 F20101130_AABXDC fickett_f_Page_040.jpg
28e0b0b3126b1d50f7e4eaef9467f8fc
12339525dae2f626ddcbe6326e930ef1f1f2fcd5
64893 F20101130_AABXCO fickett_f_Page_014.jpg
f209f443f8da0d29a21345568027f39e
15e014f88c871c9e89139ed62268f159b8eebd33
F20101130_AABWXI fickett_f_Page_082.tif
bc33764265536e89cbdd30fceecfabae
2250dd4233da5b2f9b69b7b8778f1004b69f3315
4130 F20101130_AABWWU fickett_f_Page_127thm.jpg
8fce6e2fa471ef41242d043f9aaf91ba
da0508d0172017bc8ff0e2184b829bd0b4a08a3e
29439 F20101130_AABXDD fickett_f_Page_042.jpg
5790d0f5e14db341d0828f5f803dc6f3
e128c00f21b1992e282c29ed145294820bca2453
57878 F20101130_AABXCP fickett_f_Page_016.jpg
011a70e33c85a89ca39a5f4f22507607
98da1bf95ab72002cfa72ee7ce4ab63cad04365e
69599 F20101130_AABWXJ fickett_f_Page_055.jp2
a6e296da0543174dd6c7f1172c066e24
ea52c8c0fa9fef79234dc57cae80754b3bd8f353
52970 F20101130_AABWWV fickett_f_Page_034.pro
b5c7c28baf98e56ce839a21f028c359f
95ca277ec691fe6c910bc45fb32387123f3e8adb
26425 F20101130_AABXDE fickett_f_Page_047.jpg
954d9805d769301899da918d31ada1e7
66e50fb8193c19855b69077a5a7f062860e473c4
61749 F20101130_AABXCQ fickett_f_Page_017.jpg
804046cc1caf4e9424939bf772e53d09
7fb050f306ad76a2c329cfd836db441abe540411
213 F20101130_AABWXK fickett_f_Page_003.txt
32a8e938518105001ad00a032ef24eeb
2f557a8bafe187332f0a910932dbcb9878f4992a
52363 F20101130_AABWWW fickett_f_Page_070.pro
e6b732c70d64fb6999fd89c4869e1d0b
260a73716f5404f1547a176fdda3cb08e8e5866f
32476 F20101130_AABXDF fickett_f_Page_049.jpg
269672c169eae15b710d12eaa5791410
2b4df7f17cb8a97c11a8d9f8655e2dc91afbabed
72484 F20101130_AABXCR fickett_f_Page_019.jpg
cdf73f6fabc862c3a2ed7512ceae8adf
632113b21c0eb37987c56778e01e2295b29415d6
1986 F20101130_AABWXL fickett_f_Page_085.txt
666b487ce117d068a52e17389974e859
02e2bca924cf28a2d826fa2a460823467ee468fb
555 F20101130_AABWWX fickett_f_Page_101.txt
d7b3b95eceb35a602f767f4e8330657b
6e46cfd82c6665902189886b76252be97954ad56
34085 F20101130_AABXDG fickett_f_Page_050.jpg
999d57ebd22fa2f4286c6771eac78ad4
e2e9100f827740d90cbe6088cce284f6f7b981e6
86619 F20101130_AABWYA fickett_f_Page_053.jp2
5f42a78f91cc22983215f851e9816de3
dd28759281be77ab496064b92f3c3545f89efc7e
72417 F20101130_AABXCS fickett_f_Page_021.jpg
7c690ac61480bcfcc7a6ab7f7bfdb217
433893d9290801a1bf1f6b31e922bcd0e7a42d24
2687 F20101130_AABWXM fickett_f_Page_141.txt
29f34115f60602f2bee5ccc2def891cd
64eda295440ace42f29fec463e0b7bcd37f21d6a
16228 F20101130_AABWWY fickett_f_Page_104.pro
988c7f7490f939ac85a076b25ee557db
aebad19bdce02e9fc491fd336f50afeaa2fe7abf
30883 F20101130_AABXDH fickett_f_Page_051.jpg
d92eeac77b0abcbea3562a54fe5fd16e
d935698cc4b01a29b068b77fe23ff2c21e38c398
6290 F20101130_AABWYB fickett_f_Page_067thm.jpg
6be6a7cef6941e02a496a593d194877d
5caf0fb0b3bf6a4e4210d2fcfeedf8c30ceed9e4
68770 F20101130_AABXCT fickett_f_Page_022.jpg
d39fe9676efc7b200c2b72687dfa3bbf
f2a819908dd6da633581ea7a416547e10c066a1a
F20101130_AABWXN fickett_f_Page_083.tif
4fd6e2c650385e03dbde14dfe258790e
47b1d8221b4171a1e3e4e3a379a160d822b135bd
6673 F20101130_AABWWZ fickett_f_Page_031thm.jpg
80d7f0a0b76e81dcd6940db8dd0b97be
8ab7cdf93249f37c08abdcb30340c21edb7b64c1
30179 F20101130_AABXDI fickett_f_Page_052.jpg
b256fcf3c75348cdb8bd397fb5e5ba80
21aeec320a0e0c844567350fda79e5b4f385b4bc
47676 F20101130_AABWYC fickett_f_Page_064.pro
d153083fd8e293c2ce4617f4fb30d8d7
9fd82c62f8bb0d836f4af6925a36ed7b5bd34441
F20101130_AABWXO fickett_f_Page_003.tif
6028c055a3f935939b1e5bb5c8ba275a
2a1abd37dbdd24fe794535243f3103fa69200e78
31771 F20101130_AABXDJ fickett_f_Page_053.jpg
9a4d61a157208400c4b347c20a52b6be
bc62da075c472948687656f4f9bc55bcbdc36559
136001 F20101130_AABWYD fickett_f_Page_140.jp2
eb6e9fd9b40395b6bef70650d9e74ccd
ce38509728a10a7226ce9862d3cf5ef65d8330e9
72231 F20101130_AABXCU fickett_f_Page_024.jpg
3df5df4d959f7cd76412c86bd3dfe812
683d4d0765a43975f5443cc81ff3624f65d618c9
23559 F20101130_AABWXP fickett_f_Page_020.QC.jpg
b2b5c10858738a2c4ecc9bc507a1eb1a
fcbdb555662a7ef9859b7df63f95d996e7d26c97
25318 F20101130_AABXDK fickett_f_Page_054.jpg
207725964e7cd88fc3c922cd6541ba9c
232bb4b8bb56fcb9ca6db1260bc64f4873547fb6
73646 F20101130_AABXCV fickett_f_Page_025.jpg
3ac8ae54a51bba194667fcc99e1d7ce6
5a06e74fe63a1ef266f906aead5e6c62bd1de5b8
6906 F20101130_AABWXQ fickett_f_Page_143thm.jpg
a0195f2f47205b58e54f9227143fd96e
ba5df44fc7e0d4806cd5b734aec589484c464f16
21427 F20101130_AABXDL fickett_f_Page_056.jpg
1c2cdf412022d53afb47621c04cf0c9f
54e0470b3d0eff82cc46ab254f21055ef6b96c11
6575 F20101130_AABWYE fickett_f_Page_088thm.jpg
bc6c7a3c5bbb1e43e4e7273ced390c43
2d23a4ea68dd3c9181f229c32ab3750107f86cea
66104 F20101130_AABXCW fickett_f_Page_026.jpg
7fe019df4fee5d4798105b458aedf878
9d47a2fb8c1db3d385f15d277bb80050bf10d406
12953 F20101130_AABWXR fickett_f_Page_107.pro
dc1fb60441357c0e0fcce51f060ea43e
5bc7f123d1dce897b304e2cf3327b21030c55514
73958 F20101130_AABXEA fickett_f_Page_089.jpg
9c14c5dd6b860e7fc8b6272d71c7dda1
7c0821411cfd024ad8481f92cc83e7c738b16b87
65253 F20101130_AABXDM fickett_f_Page_058.jpg
f2c7d8a9775fc291c4c786bf9954fe0c
421600983b084abfffc19425ec46f59932779489
32553 F20101130_AABWYF fickett_f_Page_043.jpg
d391a879a4584afa5f41b4be9c17ab38
047644402d8a675f93cf03c8bd5357e1819de078
71777 F20101130_AABXCX fickett_f_Page_027.jpg
091172ba8a02fb42a77cbb1a09c5f2a7
ba389992bdaa7b8ddbb3f55ac6a565dc63d94705
669 F20101130_AABWXS fickett_f_Page_125.txt
e42dab5a85fbe156c38f1453f0524952
f672296dc6563ca4a088c4a1469439f0d09afd0a
73870 F20101130_AABXEB fickett_f_Page_090.jpg
6d92b5122735581f7c21b59a29ae1f4d
0d4b5a04479a34ffcb8cbd64dd176a3c8af71547
71913 F20101130_AABXDN fickett_f_Page_059.jpg
6e6f92e0ac9c9cef1958c3b32c797f3a
bbc5d1ba7ca9d894477c378107cad783feeef0d0
73137 F20101130_AABWYG fickett_f_Page_035.jpg
437e46414affe98a965181d14c8affec
f49c64ab02748eede2b178a30454444fa7d7abf5
72411 F20101130_AABXCY fickett_f_Page_032.jpg
213af7c3011419de38212e2be5feb4c2
b8e4fab154a9e28654e7237e6cb3547758c3ad49
6608 F20101130_AABWXT fickett_f_Page_085thm.jpg
6155c5a1f6498f002d8266725fac95a4
75886a631cfe9fcc51f0e034d8c574b4fc71ef2e
51550 F20101130_AABXEC fickett_f_Page_091.jpg
c8925c25467eedb23baf717b2690b949
d8e7673065de8c9e2a1d1e6234423612a3a83c08
105669 F20101130_AABWYH fickett_f_Page_066.jp2
465125c23dc74d3cba7bffc85a97f6bb
2248e38c19c4a78ae5c54e4127b1a30adff3acf7
75475 F20101130_AABXCZ fickett_f_Page_034.jpg
fe9d93d8c106a2042bd54d3310d58fba
c87864417d77087abe0452064adadc29d8bfce7b
7012 F20101130_AABWXU fickett_f_Page_139thm.jpg
58101ed815a556f6f2eb5b6d38ebd15f
11433254c4a4dd9b740d30aa7ab066ff1cf00092
50170 F20101130_AABXED fickett_f_Page_092.jpg
06592c7b5aa0bae15e807294b68b5b69
161453da40aee81232ebe6bae2174941fb86aa84
71364 F20101130_AABXDO fickett_f_Page_061.jpg
2a9f3418c0660009b0e67a251d124af8
d25a3ce22573518bd13fda24e2355bd5da0d1ee5
25317 F20101130_AABWYI fickett_f_Page_135.QC.jpg
6b478a7e86bc87533e270e189a75164f
9610c028930fd5fad0dc14db17c618a4ab270868
82190 F20101130_AABWXV fickett_f_Page_097.jp2
5a970efd2ff2116a179545e79acef972
e2699164e6d5b2b9b15bf8879c2375a79fd4c383
43783 F20101130_AABXEE fickett_f_Page_094.jpg
ff4a923bedd49e1b586bda8a2be8e845
f8709db60a1cdd2796b4790505136b1b9fb8ad1c
69884 F20101130_AABXDP fickett_f_Page_062.jpg
de9e8caec61b0159a6b8e258771c98da
ce4f405abcc5119bcd46163a392ad8a14576b2c2
6326 F20101130_AABWYJ fickett_f_Page_078thm.jpg
4154595075067d2e4972f8c27e63d2a1
477a39c93c44c075450eb7d9fb11277b8929ebec
1663 F20101130_AABWXW fickett_f_Page_003thm.jpg
3d9e7d50151f54c66375138847f0cb33
1e7624458bc5288acaa20e41461497aa2124602a
29383 F20101130_AABXEF fickett_f_Page_098.jpg
431271c47bb2e008b541a0a19488c91d
219bbad0b82f821e81d9422c3aaa37693a836acc
70312 F20101130_AABXDQ fickett_f_Page_066.jpg
a8a2bbafb3d130fb3732db9e94bb86f1
5e5c0ab6c073875e8f2547a01a52c37e7dd349f4
F20101130_AABWYK fickett_f_Page_034.tif
2d56bc2f026c45ecbcf218986307a06c
47e80429b0522761b1921a169f03ee293f904f4a
2801 F20101130_AABWXX fickett_f_Page_046.txt
6c7926e748492f63d3b210ab0083506e
0ab597c39b9576a837696d1693fc4a9b153d1b4c
30419 F20101130_AABXEG fickett_f_Page_099.jpg
b94c60c1d55767fa1ae9d154d82f793e
e81fe209dc92bedb1ff4896a1418d2b9c52aa5d4
72398 F20101130_AABWZA fickett_f_Page_036.jpg
b6d7dedc1eb18493058798eeb8dfe3b7
621aa0c3745ef11cc31a31aed57e487f37fb7f81
69374 F20101130_AABXDR fickett_f_Page_067.jpg
dc1624929316a12e9424cec3d0347156
6c367bcfe7b809a0c3a78160959ef0b0f25972bb
22830 F20101130_AABWYL fickett_f_Page_015.QC.jpg
7feb7f7a34d4f49c7ce8fb5677cd8eec
f4a2b965aa232ed07be56350b5653407e830453f
F20101130_AABWXY fickett_f_Page_146.tif
e5a30300c53b03f0db9055af1848700b
99d7161f4614f0ad8fb67220ba5df2353cbdeb25
29867 F20101130_AABXEH fickett_f_Page_101.jpg
3132fbee60dbc342b97e3650426f3bd5
847e0723eb8323be8b7c319286c6f75c50479bc1
754794 F20101130_AABWZB fickett_f_Page_102.jp2
d1be8af199d9ccdf259e03b7fb9d062c
d4a69cc0b20d6570045fd78313b206c65b481941
74659 F20101130_AABXDS fickett_f_Page_069.jpg
a236034a33f3ceb63e1bca93285b6159
38e3860a59ad0d9725984273ea27d514ce9e4cc2
1044 F20101130_AABWYM fickett_f_Page_127.txt
97e8bf5fb36ff2bd84a865c7c57fa2b7
db99504a5b226bd0222bbca0d0b4669ab2bc54e5
5006 F20101130_AABWXZ fickett_f_Page_102thm.jpg
5c29c179d3b9cb581cfdd5fc0a0a16fb
c720af9c54a53ee5a8c64c0a0f25eba4efd00e48
54407 F20101130_AABXEI fickett_f_Page_102.jpg
9ce0cb5df6c79afbd92ccd2b4f99715e
1f357fc19c0fabb7583630376db04aa904cdcb9b
37297 F20101130_AABWZC fickett_f_Page_006.pro
1b1b0d4b280584550a04da87b2771ab8
a73c7af00be3b56596d0eae827657161b5a5daf3
74771 F20101130_AABXDT fickett_f_Page_070.jpg
64f4fa92aff30ac5246ce9a6fc5f0a7d
08373cb895a64ac4ec6825362578102b1fcfe480
4782 F20101130_AABWYN fickett_f_Page_005thm.jpg
00cdcd45e74ea9df2ffc4d65dfad0af3
e352d04d6911986b5a7cbfa173a185c45a470116
40385 F20101130_AABXEJ fickett_f_Page_105.jpg
b6562f6d9fd1956afea1c8d1cba2544d
8d437bbd7d8ae4cd936baee705212e1009fe4bf2
F20101130_AABWZD fickett_f_Page_145.tif
2fda33094313613342ca348bc431d0a2
3906b0dda9f10451fb8cb222b843e8ec6a34c247
72485 F20101130_AABXDU fickett_f_Page_074.jpg
20a2640eefd41c0ad52badbe6ca87b9f
697ee4bc4b448eacbea2450eed1984d9f08a59b0
50735 F20101130_AABWYO fickett_f_Page_056.jp2
ff67b00b43c87d809dfdf163e37c2859
ba0712b507a2d48dec79c1aee420f9439efeb1e6
49238 F20101130_AABXEK fickett_f_Page_106.jpg
caf61e5b5532be7fc0415f7272e8b369
34c6d7078bc8400646b4630321c9298d132d1ce4
8282 F20101130_AABWZE fickett_f_Page_047.QC.jpg
63985df25cf13d94e9c4a7fc9034b0dd
49e418c7b64d857a024c701e3bba2db8463c8a0c
71375 F20101130_AABXDV fickett_f_Page_076.jpg
f577a2fb95124230491947375e98baec
19092a80ff79f7aed97b4e96c4db3b90cd4a785d
72415 F20101130_AABWYP fickett_f_Page_080.jpg
25be21c5ee8f56fc6788faaa7cd4f21e
93436ace687598e247c844c1d6b7dfa6376cd509
36473 F20101130_AABXEL fickett_f_Page_107.jpg
ff0414fb65d3b6c9497aa8a2e07b38c0
bb84c7e0f19e6e2dbe8130f263fb99beed0900d2
75260 F20101130_AABXDW fickett_f_Page_081.jpg
c3b7ceeb222e91633aeebb9c642c20b0
cb60b3038e0824ebdb7198da2d0dc93d778887f2
1906 F20101130_AABWYQ fickett_f_Page_068.txt
8b7bc79b212013666d213a7968f0e185
190a098a5caab8376908b29fd21998829e0557c5
32385 F20101130_AABXEM fickett_f_Page_108.jpg
583c6a67f6dec338d83986cb8e529741
bcf9cf54e608ea6f4925f5ec9bcf6e8e17192a62
3197 F20101130_AABWZF fickett_f_Page_051.txt
279a4f9269a65619ee1f4fa569d2baff
ebe90fc0aa30fac4108387480e6006789a0771be
71988 F20101130_AABXDX fickett_f_Page_083.jpg
9651406d9d9bbbe4fb164019bb7b3816
bbcbb657cb0cbbfed3ff24295d62261827e799e6
110507 F20101130_AABWYR fickett_f_Page_085.jp2
d19c4dce74c470032e9e3de12b414db8
09e570434608f6f8eef92878d06749254549d782
27653 F20101130_AABXFA fickett_f_Page_129.jpg
e2e5300066ab984c740e8fa15c591fb5
db50e6238b490d57a74cf64e0ba6d085aaa1b69c
51361 F20101130_AABXEN fickett_f_Page_109.jpg
bb835623f86fd29cf16dc59feea17583
4a703dc56fa75eba2a0d65aacd5e2fd1101eab0a
1983 F20101130_AABWZG fickett_f_Page_076.txt
2d70614a790adee7e78c46ef9b1203dd
69088d8f7b8c81c551280694d1cd581b790cf139
74294 F20101130_AABXDY fickett_f_Page_085.jpg
02bb4f83be81636f15fe105bc6988691
09c71d9feab188b758603580a9901eff41c2dd65
2933 F20101130_AABWYS fickett_f_Page_099thm.jpg
cfc8c568564306ad39dd5962219c05a0
2e27ce8f20802bd748129f7065911921f17f8c36
75551 F20101130_AABXFB fickett_f_Page_134.jpg
5727045468abad6ccd86f6f633689240
e236d994f1dd9d105998ee8e140c7ae13387ba5f
52216 F20101130_AABXEO fickett_f_Page_110.jpg
f5a98ad2cfc5adbf47cdec0da29bfeeb
3e2add400f0441f0fd67844f2790e2bbe75500ec
11371 F20101130_AABWZH fickett_f_Page_100.QC.jpg
fa892e465fb80ffd65e3f276d1415710
a1994e2b2738f33e2905ffc48e0827506f773cce
73959 F20101130_AABXDZ fickett_f_Page_086.jpg
04f1289c37c07fd065e9cb7d0881b058
32e075615f91993719c76800480cc7f5c89dcadc
2551 F20101130_AABWYT fickett_f_Page_054.txt
e7965906073011b72bc45c12dd47f723
d3b38114468506274caf1406ffe9e78f812f5e21
83865 F20101130_AABXFC fickett_f_Page_136.jpg
726b593a82a04d5afbacfab96b1cda57
0110dda1a14f9d3a52411c52c95fe86557fabbef
65331 F20101130_AABWZI fickett_f_Page_043.pro
89ce558adbb357bd23b184003678f71b
a66a6446b5908159cf10105daa42cc0efd717e0b
3217 F20101130_AABWYU fickett_f_Page_050thm.jpg
b97427b66c38a591b93a382f46a4e8db
4245cbb9a996833fe65aba801bf654a8dd7f40a6
89805 F20101130_AABXFD fickett_f_Page_137.jpg
9a752e97f1fe8d29b08a6fcd2ac83fca
0474215f85069082dace84bac5ce7c6b44647e07
53095 F20101130_AABXEP fickett_f_Page_111.jpg
9f799dfb127b6671ac90156141456429
31264e4a4a6a8927715700e480abcd857892dbbf
131276 F20101130_AABWZJ fickett_f_Page_138.jp2
7a1bc9caeb898399bfcfb661fd45e4b8
c355155a95e04e9f329037927bf51b3abaa95a4d
2063 F20101130_AABWYV fickett_f_Page_031.txt
882764d8af6026c568f7852f71cb94b1
a2da2e345127a5392d7f2dbfbeb4fa5cb740abe8
91272 F20101130_AABXFE fickett_f_Page_138.jpg
160d3d6ea2fa5e4ec3a620d4b60211c7
b58b1e8685ecb63b89c282f3a29eb7eeaabf5355
55749 F20101130_AABXEQ fickett_f_Page_114.jpg
eca9f12f1265a42605e13f0d5bf598b9
2417b2fe69e53d0745db69cee320f6d4d206af07
2657 F20101130_AABWZK fickett_f_Page_143.txt
5a6befa809fc17fa34e5b2eb14181b88
c8d13497555da8f57abe10d0d31bceb49d7cdead
34090 F20101130_AABWYW fickett_f_Page_056.pro
95c3da8e9d39940369a3bb37c54694d8
071e7a174e0b4540c406b2e2afc749b18d544005
89284 F20101130_AABXFF fickett_f_Page_141.jpg
91b340ae592fd848c1e7278fb2fb8625
2d858e3f679c2f320290f9ec8db0bf928427448e
59410 F20101130_AABXER fickett_f_Page_116.jpg
bb7d9315d302b09457759fc4de2484da
621ac2f91ed821ac5ed01d871a4fe70785c54f0b
101841 F20101130_AABWZL fickett_f_Page_029.jp2
404feece2d9dde7876476e7199ddc73e
713d67ccb94c102b27b83829f081ba5cd943514e
6448 F20101130_AABWYX fickett_f_Page_136thm.jpg
9a84ac83e771b11b472ec34752e8dcf8
e9d9e71d7932ed187ba92f1f24bfbf462f1ddfb1
90269 F20101130_AABXFG fickett_f_Page_143.jpg
bfc100d743ad191ee319031cce7f26d5
8a1f42a7ff9295ca2f052978bc3460beb4098915
36793 F20101130_AABXES fickett_f_Page_119.jpg
a758a3b23d7c47f48f9fff794970f167
2a7d2a82c68c8d88c32953db9004700123a354e5
23129 F20101130_AABWZM fickett_f_Page_027.QC.jpg
3c21fafea26d266c19126db3a855163e
e443a1c504e753ce93524ac7604d28f79e4cd6b3
51067 F20101130_AABWYY fickett_f_Page_082.pro
03d8ef6cdd7f3c888370c1fd7e4df127
6cc9fcabb47199fb0d7c77c24bd17c75ae7e4f4e
91534 F20101130_AABXFH fickett_f_Page_144.jpg
466f73491b1adc592d54d4baff16c0ff
5e7e7d1621451fa90838c85782c798f3387e2ea5
59140 F20101130_AABXET fickett_f_Page_120.jpg
3c11b8b614b873ea2567d8a2938b9698
9f510f3baa9977d666acdf1def26e3878c8d5a24
72719 F20101130_AABWZN fickett_f_Page_063.jpg
5b2ddafb56c2468648500431ada3c3f8
3b620649f28fe2fa3eccfaaa6e3f959872e7b1dc
18126 F20101130_AABWYZ fickett_f_Page_102.pro
cb2d48e39257cc684bb7503c410fed19
77b358a9c4c0fadd8b5546dede8d6cb57044b118
17735 F20101130_AABXFI fickett_f_Page_145.jpg
8b6b0c5e25fcd56a1da95df1313f57dc
c723a8815a3cd844cf9a972a48bb239ec8242b9b
36732 F20101130_AABXEU fickett_f_Page_121.jpg
ed41196a941f10accb9e72ff0e173730
b2dfbc6751ab1263b406ffb500b2580136093d62
50689 F20101130_AABWZO fickett_f_Page_085.pro
eb6928e52cf5e2febd0481394e4b1087
da7f1c18d8014bf8658f4bd9e34db1751b9463dc
26498 F20101130_AABXFJ fickett_f_Page_146.jpg
47284a32147d40b508d0aca4a020908e
a09695c6df5a9c582a062f38074b39add824ec3c
31804 F20101130_AABXEV fickett_f_Page_122.jpg
d81320340d87e7e535c9968caec2f6d5
dc19315cab02eb79a2be2769f51933568c2498e8
23739 F20101130_AABWZP fickett_f_Page_080.QC.jpg
44a3d615f04f15323bfbf79839046a6e
68bd7cdc59294a2fe9f7becd61af93d9fb8bbda0
28005 F20101130_AABXFK fickett_f_Page_001.jp2
c14c6fd5796b6d8921bc9f5ac79a8c19
ca1f3ef0b8679fce28b9d4b6153b44afaa8f8469
30448 F20101130_AABXEW fickett_f_Page_123.jpg
8a349333627c2d77aa40e1ba494a2173
b579a8e978d407cdc61ff0a7f07af579d1391603
678 F20101130_AABWZQ fickett_f_Page_123.txt
863577a10379c0719dcdf07b6e36d3a4
9baa48050f23232f6030b18634a3b992ff90d0e7
5544 F20101130_AABXFL fickett_f_Page_002.jp2
22eab06f78f8a7f2256cd7e5197f5078
47e05ef4fc22987f1cca8745768e5fb02eaa830c
30043 F20101130_AABXEX fickett_f_Page_124.jpg
2c23ceab921575389b09bb0ef0e2ce03
2e04b3d477aaa5bdcf95ac200f36e3434d3a46dd
59103 F20101130_AABWZR fickett_f_Page_136.pro
3bcf9b0fe380fb27432d8bdd87e8a44e
aa47687a95e9d5a6ccd1566dadb45ee21fbe0b02
110805 F20101130_AABXGA fickett_f_Page_030.jp2
74c7b83dc7a40dd18cfef0ce281ea6cd
6ec1933da8d18fd88169431dc8ee489723c8b24b
962414 F20101130_AABXFM fickett_f_Page_004.jp2
1e32d87aed47e90ac314e4db3ada4508
4fd8bfcc71f2818e3edae44366ab66640b40fc2a
30541 F20101130_AABXEY fickett_f_Page_125.jpg
bb3e3ac3b295dccde6e957562f3fb90c
423d337ab87052c4abe6afd21b90db56dd9c392c
14584 F20101130_AABWZS fickett_f_Page_092.QC.jpg
7e1bc800fa9c833de83aad8bb8160fde
25e976bf81af2276e46b61f8ad38ed44cb50426b
113234 F20101130_AABXGB fickett_f_Page_031.jp2
09013dc810b9da3f43a9913d619c90cd
552b5d41e8bb0a6d8730e1ce9d2d37034b429597
854918 F20101130_AABXFN fickett_f_Page_006.jp2
6815c80d46ceb37f47cd63fb65939d5c
253bc1f63fe1f9201e34389bae9efaca8e3cb882
41869 F20101130_AABXEZ fickett_f_Page_127.jpg
4509cf2ef4c74859b5a98c7d60013311
2fa322699c3ef8bb5098b84494de494c86812799
9438 F20101130_AABWZT fickett_f_Page_101.QC.jpg
64ebc4fbcd667b7c4b203070467e2170
ffcecb1096d3354b45512d4c2498a59bdb27fd15
108804 F20101130_AABXGC fickett_f_Page_032.jp2
135e021d9ec87c9636aaa3355ad0347a
32d896b86f019c981582a3f24e38fdb0216dcc39
1051983 F20101130_AABXFO fickett_f_Page_007.jp2
b3535318daf2ea0f667e8be9b6e01ea9
8946cacdf2d9f6c7c387fc1775435b58e948706a
37306 F20101130_AABWZU fickett_f_Page_012.pro
fca08191cd516722545cf0cb61ef0277
3912b32fbff49338567183d06f5e8991a2b81bc7
114258 F20101130_AABXGD fickett_f_Page_034.jp2
33af2fcdd7600d2048013e1299529523
3586a145855abf5d6ffae219e498f69c703f6c58
1051890 F20101130_AABXFP fickett_f_Page_009.jp2
0426f4c8bd4d317ba3607a1c130bc998
088dd3331e79d3438b565c3fc86cdf148c18d04f
18674 F20101130_AABWZV fickett_f_Page_012.QC.jpg
6ee4adb99b01746da653ccc0881ee8b1
cf61b37886671d72831d0851d6270f9e62d093a0
109223 F20101130_AABXGE fickett_f_Page_035.jp2
95b2b6ef0df0e2ad3ac6aa256055b1c6
b155f955924b3053d9d1b3615e9a0e0d49a16c16
10482 F20101130_AABWZW fickett_f_Page_126.QC.jpg
383f408770c3d026c8f8a753169072dd
02a501e960e2fe8bc8c235ddd8682ae35352d1be
111556 F20101130_AABXGF fickett_f_Page_036.jp2
9d4cefa2c6cb8692d8cbe5de12a42eca
bc091e7885dcf518899864b7a27f396f3322b0dc
1051938 F20101130_AABXFQ fickett_f_Page_010.jp2
0e46f53c2a1a4e32b480b00243251857
5402f545151c33ca8b302bfb9a07e243844065e2
F20101130_AABWZX fickett_f_Page_120.tif
495dc0b709e6db4d0bb284972e4c4c70
00d3cca17d2b6ff57c61da2b47c493cbdcd2cf0a
105603 F20101130_AABXGG fickett_f_Page_038.jp2
1d504174f3b86d2973d86afbd06e0cc4
d83685ecb5fc72ce86306d948d52f8c776b1488e
83994 F20101130_AABXFR fickett_f_Page_012.jp2
770e27e60fdea466c8d214449766b163
2e8815642b19d45912ed81a6de8de3613f0b6f82
F20101130_AABWZY fickett_f_Page_126.tif
0e862f5fbf391b8242266ea0796dd616
527337fd24e9a3925c6656773ffe679fe2a3f067
75365 F20101130_AABXGH fickett_f_Page_041.jp2
6ff72c6113de1a0a4fa228e517b6b1b4
0f45824ee4d8f6dd0e70af0ce02a8453ec628abe
96891 F20101130_AABXFS fickett_f_Page_014.jp2
852e6b70e6b6cf73a280138d00b96f18
146adbc5ae7df37e3c7e72723a4323778788701f
42609 F20101130_AABWZZ fickett_f_Page_039.jpg
66badd1ea69321a0cf11ec35319ca66f
0d798617c54e24685050ad36b57867b5f242a662
86921 F20101130_AABXGI fickett_f_Page_043.jp2
0492d6863cbf587d594ec6f3ec1b180a
60028ef3fadb3856b8da057567f06560a4460d39
86163 F20101130_AABXFT fickett_f_Page_016.jp2
0658ccec1811708e9f4c187bc0188191
e79182a0e7b179576753fefa905c53b7b2cd6d04
75198 F20101130_AABXGJ fickett_f_Page_044.jp2
97fe81591b5caee7dd25bd2f6cb2acdf
51dd213143977b2ec2038418e2b9d8c7a08f5068
89180 F20101130_AABXFU fickett_f_Page_017.jp2
0891c9c68269047f49a5cc92735008c7
366c7e4f3339d1bc8e9224ba4fc6adab2c789f8c
76946 F20101130_AABXGK fickett_f_Page_045.jp2
1d364ecd01c75ee83e2a7d52512fd429
d35b6c4f1478faded60824fd8b4e92e38890206f
109407 F20101130_AABXFV fickett_f_Page_018.jp2
3feee8c216381e5ed62851d4ee79922d
85a46b1197f18830bdcd83f75e415e0674cfc005
75818 F20101130_AABXGL fickett_f_Page_048.jp2
93f20521b0ed0d76b8e1d6f3352246b8
34439adbfc1e3363eeef6d7f4b7844407a0ea5a6
109364 F20101130_AABXFW fickett_f_Page_019.jp2
de670212956cf5688d2301c47f7bd36c
f572625476d69a44f426c738e3033df26953aab9
106641 F20101130_AABXHA fickett_f_Page_083.jp2
aad17ab9fd6c6f58e1ce57033e928eba
5ff8a345cf083aed8a7a25809363bf40130b8a9b
92013 F20101130_AABXGM fickett_f_Page_050.jp2
e125ce846fca1ebe3d711952b875f966
76cc4b88978046c4f92557bc7889bf10b9347e4b
109959 F20101130_AABXFX fickett_f_Page_024.jp2
7347dad57f22be065d7fc14f182d80bb
9c1db808d4113c9d0f254770902a123442239eec
112279 F20101130_AABXHB fickett_f_Page_086.jp2
784bc9033928a11888bb11fe5a0988b5
c55422ae034c6ea7c64542f83abab265ccf6abe1
112358 F20101130_AABXGN fickett_f_Page_060.jp2
28eb02ce1135f84b09011e60fe473d77
0d63ad60344244cce4664d4cfa0d0f8c3ffb7058
110298 F20101130_AABXFY fickett_f_Page_025.jp2
6ee014c5f94a8bcbc2e937a3e9df5ce6
b9ebea44875cd4d08031a3819737e29b87101ff5
109340 F20101130_AABXHC fickett_f_Page_087.jp2
03097fe7a6932962781302b3b018391e
70ddcf27f15a99e9683ed2bc397c62bca58efafe
109499 F20101130_AABXGO fickett_f_Page_061.jp2
8d69c3148f60cb8b9d289c6a26972a50
c8e71526d58b0ee4653f656707eb98d0f58d79a3
110139 F20101130_AABXFZ fickett_f_Page_027.jp2
66ef6f599db04ab1d58debd9623ae14a
fb09350d28d0aa54b243fa6060c39de22efa0e73
106200 F20101130_AABXHD fickett_f_Page_088.jp2
91a32af4b49ab3dcc3e1f2cdb960c6ce
61e4dd3ceea000f2d9136b1eb7413b99e8afd91a
105087 F20101130_AABXGP fickett_f_Page_062.jp2
f8cc657ecc5fb085bd1b08c352409fbf
b7ed3ab2cf3d72b1987499287cfd3b2ac2ddec69
110247 F20101130_AABXHE fickett_f_Page_090.jp2
d63e6cd38de8db1aa41d986b3cdff0f8
bdff613aed8a4fc0091171245fd3710aab4ba853
107374 F20101130_AABXGQ fickett_f_Page_064.jp2
b4b30085c395ae37da012285436d9be3
2fdd97356c908d729d8ff2fd7b5f8f63df967ce6
70521 F20101130_AABXHF fickett_f_Page_092.jp2
c4e42866e7ced36e506fae54f2e1075f
4d0ec4c5afbbb5b2251d91b711a49bf9ae48bd49
85350 F20101130_AABXHG fickett_f_Page_093.jp2
5f98b515a0e82afc8432c088fc52c81d
3083a9816b394daae6545638f4ab652a9f55defb
106043 F20101130_AABXGR fickett_f_Page_068.jp2
e6e9b91e12b1618876a9001416b38255
c4593653e487015e0ae3ddd526ce66410d8ec72b
98677 F20101130_AABXHH fickett_f_Page_095.jp2
4c513db525d9d9734942411a5cb702cd
a8a7f44ea1f83a88ea02c181ab085528e6053388
104996 F20101130_AABXGS fickett_f_Page_071.jp2
dcd5e028815a5457713b7976400c76a9
8d9afd06f1689a58ca1787c6ca19de2f8c1e3b64
544663 F20101130_AABXHI fickett_f_Page_104.jp2
af67da85d7aed23c516849cc3e192617
871410cffe8e34fab24b4db6dfaf5fb4cb5fdc7d
108362 F20101130_AABXGT fickett_f_Page_072.jp2
12f9b4a925e7bece239de40ca2e77cce
b96e3da18b84a260c309bcba34e8855a0de37e05
639138 F20101130_AABXHJ fickett_f_Page_106.jp2
881f4ae6f9381700266dbdd89f4ef96c
60f1d3541033b2424977b31775d89989b2764e69
110346 F20101130_AABXGU fickett_f_Page_075.jp2
f728d2b39936b50536b7265cb1cf3da5
51425579605a9bcf47fd38a1aa72e5d532e03f58
464647 F20101130_AABXHK fickett_f_Page_107.jp2
cbeb2704f6c582695cb22469a603f47f
ea3ddb95c0443a38aa71861e7ec4e7aa31957f9e
109839 F20101130_AABXGV fickett_f_Page_076.jp2
3c4408467b5696f943eb5977e1eba674
839df9d20052d7801881eaef7c161725ccbac8a8
412264 F20101130_AABXHL fickett_f_Page_108.jp2
922829b845e61442ffadaadceecc834c
863730baba40d394f4d07b3f4fbf2cc71e035200
104496 F20101130_AABXGW fickett_f_Page_078.jp2
0b5b57666d440eeac340556176c0d166
20b64259a31441a28f506ca15ed63a4b5924e420
666924 F20101130_AABXHM fickett_f_Page_109.jp2
ed4a9254d30896f1eac2d222f3ecb0c7
f1ff871adddd1d64450ba719bfd21b85648ec824
108839 F20101130_AABXGX fickett_f_Page_080.jp2
c0043e31cf2c3bed4ffc4f2121ef6519
3af1088cb4b7e503648bef1009022cad3b075747
518678 F20101130_AABXIA fickett_f_Page_127.jp2
85246d2405b3e7ce47e91974dabe87b1
925e8f6866017100d7738afc78e8ea51e1e6a4d0
693784 F20101130_AABXHN fickett_f_Page_110.jp2
e7124a3da7b9274049c2c2d447300669
38be4a4aa6b508ebb8fb6ce30a01b0f4ff2e3b38
111629 F20101130_AABXGY fickett_f_Page_081.jp2
a4eafd7156d0aae3aa407460378be230
7e386343d599b6a090368c73484dc9802e404f1d
62310 F20101130_AABXIB fickett_f_Page_129.jp2
0d70b0d110cf9d1de3840b8a84e57063
b70281df945de121f3680a5d0cdf191effd70bd9
724375 F20101130_AABXHO fickett_f_Page_111.jp2
e0c5ce9a97ee599b260f030d9e32ec4a
18d2ccc241c6c75dcaf175ab1bfa215a206cb08b
110621 F20101130_AABXGZ fickett_f_Page_082.jp2
7a9836a5b22efdfbfbe55b050bccd186
f03351dadb9a18385aa1213fd6e458ba776038e0
54627 F20101130_AABXIC fickett_f_Page_130.jp2
7e5d3aa1d85179acd5c3977e64c2f2f9
18f45d459a9a608c528d372dfa82f4db900621e3
724131 F20101130_AABXHP fickett_f_Page_112.jp2
4cc9aa6b4c0fc05790752ed8f4428099
6d8e16706e0255dd6266c3b59c9a5e0e10dd5ddd
57165 F20101130_AABXID fickett_f_Page_131.jp2
996dfc5c484a04a0a65a502112952c33
058cf381d2818b0b5e463e82a20d5baa28218583
746318 F20101130_AABXHQ fickett_f_Page_114.jp2
52c116c87fd735529f91760d5e615968
f2d2d51f7aece08b0f108bd89d5dde1bc0d88cd9
56848 F20101130_AABXIE fickett_f_Page_133.jp2
bc3b7edf954c0584c6aea3250a2379f3
c23b0045d5d9f55a840fa0d57762eb0fec94f002
765977 F20101130_AABXHR fickett_f_Page_116.jp2
edd60eae6c190f87a111fbd64cf15f13
2bf390969ff60f484e32b4089cbab54ba74a605d
125990 F20101130_AABXIF fickett_f_Page_137.jp2
8b57a7732dbd3ca3d77d84811893064b
41f57dec85f12670bcceeb436cf3aaa1257f5bb8
128893 F20101130_AABXIG fickett_f_Page_139.jp2
f73e19e7b7c3bfca310ac550c8b7dcd8
1c0f36c3cee1249873c558c591dccb86d276c84e
133512 F20101130_AABXIH fickett_f_Page_141.jp2
caa043b0deceaf020fd003b898507fbc
b46546acc3ea1cdb14d39761e5996350e764359c
416886 F20101130_AABXHS fickett_f_Page_117.jp2
07613789972255a6e3b35a4b41dfdaeb
d963bad83d196e6c807cacb0edb5155d332fbc8a
32913 F20101130_AABXII fickett_f_Page_146.jp2
6e2db4d9d9351c3260cae528c104c168
d6af590d3d7eba81eb367ff4b1a95c7b7bb5f7ff
420001 F20101130_AABXHT fickett_f_Page_118.jp2
371993cd1a10dfa3baba7c63070b5820
017e0a87b7de3be8c795a3ba610ccb3546286514
F20101130_AABXIJ fickett_f_Page_001.tif
aeaf2cf1027d32274814135fd9bdacfb
6a3e1ef996fe2a0a0f4e8c5de3f14069bda7fd3b
449731 F20101130_AABXHU fickett_f_Page_119.jp2
8e4bf0a06f98493e097921a1a16ecc94
d349c9af80af3c48317c37b19b3a326dcfc3925d
F20101130_AABXIK fickett_f_Page_002.tif
f2d89bf9cff096b16809be219c0d2266
927bf53445771578aab92672c83cf6ed2d216ca5
817512 F20101130_AABXHV fickett_f_Page_120.jp2
744a0d29c789d340545119fb60c35422
c1933cb8a872aa05a4a652d33f3c11c02c87d148
F20101130_AABXIL fickett_f_Page_005.tif
49619c1158f246a53d9f35394af54ced
382a931603dc18d5b650234d3bd24ab704cdb6b1
359812 F20101130_AABXHW fickett_f_Page_123.jp2
d5b78981f19610370fe0a4f94e9b9b36
a78e5d1e4ede1099d7a1130e9e7e9256b193de4c
F20101130_AABXJA fickett_f_Page_030.tif
a49595208e439d7219061e648d3243bf
c9cf850e6af194034a790b23c1ff08ff4ce2f1be
F20101130_AABXIM fickett_f_Page_006.tif
4bb6e2f04f773e91298c09985ab3651f
3f2bb1f2b87f6ca091200b81892ec97613f5fe63
349344 F20101130_AABXHX fickett_f_Page_124.jp2
e32d152f079ba2a289a578b06a4ed66e
d537681f4661657cd7bae3ab0fa486ceadbc3453
F20101130_AABXJB fickett_f_Page_031.tif
8b1bf5c82a4e2fb385824b997ae02b4e
0230e079bbb84d63061d5b36d1d4ffe2fa0765ea
F20101130_AABXIN fickett_f_Page_007.tif
cbb6432a433d2c80f76b9b535f04f798
67b8ddec7dda7a22791315d78a156ffcf617bb6b
374130 F20101130_AABXHY fickett_f_Page_125.jp2
89a6ba49bb115398cac8c43b565589c8
940e764c0323ee68084c5135ad1528b81e928173
F20101130_AABXJC fickett_f_Page_032.tif
95b083e990cb67a8db4ca86dd3dc1edc
ade2197e17392082b2ff3e90bb8803bc81d3129e
F20101130_AABXIO fickett_f_Page_008.tif
4ac8423d651155973d33c6efe02041df
a8ee64f7865458a0a82a46db9dd84c59d7a7a48f
448607 F20101130_AABXHZ fickett_f_Page_126.jp2
22257d9e72a642fad80579bbcd14de57
738b3555b1dde90214fab59b11d3b7843f87355d
F20101130_AABXJD fickett_f_Page_033.tif
c123061e3ad1837fe94353c6c6c0c332
48300a96493c177c28cfedfaf0abf3dd5f6d6c6d
F20101130_AABXIP fickett_f_Page_010.tif
7614c0d30ad834cd1e199bb9bf99912b
0a0377aed38f5f2b0505e1d1f1f6e59b6f1d7137
F20101130_AABXJE fickett_f_Page_035.tif
cda035a6db965c9993b8e5a753547e82
e3ab8e5def9dab05beb8e82f3ce7a0f527581dfb
F20101130_AABXIQ fickett_f_Page_011.tif
7b2915dd61518f5964cdb1d5a0f5e0f2
99fba74635243aaa9d6b3f410892cd479a366abe
F20101130_AABXJF fickett_f_Page_038.tif
1d4eb153334e710297edcff6079dbcdb
5d0430527d509b5f97fb8d6f7f86408a7746ed78
F20101130_AABXIR fickett_f_Page_013.tif
7136736839d280a425180015ea57063d
8b6b3f2a8ec53f346e45191cda97e9fd9187a59a
F20101130_AABXJG fickett_f_Page_040.tif
bc0e1ba65a8adb892f31ddab45ae531a
5fc8bbe08d1114ba9570c361ce5329186c93ab28
F20101130_AABXIS fickett_f_Page_018.tif
e9fb02b9d22beb2d60b37b245eebe257
a06160773384ea3f9774d27b14b7eb13fdda36d0
F20101130_AABXJH fickett_f_Page_041.tif
0ee4c64b4f808bca12ea57b79a3a8049
34d2d675665da89f3c0b66e33653c74a040e0027
F20101130_AABXJI fickett_f_Page_042.tif
8683cf49fe3c627a5a89b829239dd6f5
4bbc86fc9f0a92280d49dd8ead5bd156c319f1d1
F20101130_AABXIT fickett_f_Page_019.tif
1e734851d0437d63e0e379fed67e60a8
3e8bfd2a75d6f84a768d0ec0052c149aa17f7aff
F20101130_AABXJJ fickett_f_Page_043.tif
05fc43a006ff48d8b2b394e546e42ce0
87e5ad9fb7b34884a4e79dd1f6d1998ddb6c3520
F20101130_AABXIU fickett_f_Page_020.tif
9d91a57b6c66ef193c014f5d0d61375b
eaaafd20d09be87a643e39e17e7daea82f5a86af
F20101130_AABXJK fickett_f_Page_044.tif
f1517eee6910439b105743d4692e0459
a356b5f9fde8e3056d0920031c8d310d5410bd85
F20101130_AABXIV fickett_f_Page_021.tif
e73d3a75b72ca2983523719a5ea85bca
b445da9e4532175b134530f34339cd92bcdfeb00
F20101130_AABXJL fickett_f_Page_045.tif
0af1867ef7c1f1774c1a15fa24088d99
80b0245df0764b26c64b7bc3343a93c947cfee90
F20101130_AABXIW fickett_f_Page_024.tif
2bf73faf3d4ec5ed35511fe15b0e0484
fdbf579401c5051f9fcd55837b96ad2387188cb2
F20101130_AABXJM fickett_f_Page_046.tif
5659727906c6ee766b8280d372fc53b7
8a110572b4f700464af06ee1acf9b5dbcba0f471
F20101130_AABXIX fickett_f_Page_025.tif
07f9a170079f43a8c8181e5908f09e6b
c1edd34e8d37e661362edc694989f2f436b566f6
F20101130_AABXKA fickett_f_Page_067.tif
c853613ffee9eafbab102d67521174cf
a5ec7d65395ec9399c6f76c40b53be616b67476a
F20101130_AABXJN fickett_f_Page_047.tif
2a4b8d2543ffa573c656688eeda6f4ff
a09ba83f27b15abd77b31b983b4a75ba60e1d8e6
F20101130_AABXIY fickett_f_Page_027.tif
026c7a0f49c4fd75711c059e15fa63c8
4954cf5d56989adcf9a420e20d3b2fcca3576f05
F20101130_AABXKB fickett_f_Page_068.tif
d8487ff65a381db714f399f71f2cf3ae
0130e2e08bef3d11d8c356612ac6a6fdc53ff29b
F20101130_AABXJO fickett_f_Page_048.tif
654c8fe5431f49e7a2cb1c934ace9dd2
37e45ab8bd44fcb76b0087caacb3b06aa42ed850
F20101130_AABXIZ fickett_f_Page_028.tif
b59ec068e38ff90775ba453a54fa56a8
abc008d274514b8e59b9874d695c98dde462ab3a
F20101130_AABXKC fickett_f_Page_069.tif
b0da88ad12667d4c168a07b28a295a5d
b7ca199e1db8de4e6464da57f82d2228b50f0cd2
F20101130_AABXJP fickett_f_Page_049.tif
1210c95aeadfd0d6ba4fbecf0b7e16af
8340b3aaedf13ddc7e363ad96eb052b8019b94e4
F20101130_AABXKD fickett_f_Page_070.tif
c30e34afde61f83a49af681beaea407b
5c11c542691092efe409d0097f3f06ed47b27b2d
F20101130_AABXJQ fickett_f_Page_050.tif
e08c95e115005cf294d57ef0204f7e00
5f981eb2ee5bd675ec68a1e16987910cc80e03d3
F20101130_AABXKE fickett_f_Page_071.tif
931c3fda81e13d94ae3b883faf8c0034
9142fdc2e6299d4f89df0bfc4edeb9c87ac653b2
F20101130_AABXJR fickett_f_Page_051.tif
c131d78d7725644ad307c91d61ff5216
8a978852659d07c85a8acb6e8b2784b63262ae0a
F20101130_AABXKF fickett_f_Page_072.tif
036ad8f3e48fb8c21ff3983b5c3ea25b
cfa51d132662f690f1a804e6219547b14e137a1c
F20101130_AABXJS fickett_f_Page_052.tif
ea00e95ba3b32e47c6fe46f056d274aa
9f6ca4b4fe00f1cdd48356c8ca8a7b68270d0629
F20101130_AABXKG fickett_f_Page_073.tif
7a8d2e90ae3773fb0d3827891e389d91
0c5b8ae8e23b0ddebab8d6c1df2dbc48ed741844
F20101130_AABXJT fickett_f_Page_053.tif
b9840f6f4772245653e61d2840e9a0a5
6d63f41b66082032583fef58f452da03b0d6efbe
F20101130_AABXKH fickett_f_Page_074.tif
df614800bc2719739ffcfdaefc860a7f
9b5c4bf2585fa049a6060307873554985fecc0d2
F20101130_AABXKI fickett_f_Page_075.tif
e0c59d2094ee7a4489df7d03d32bd841
5c50f845a43e96185c0ced11335b41408488b22f
F20101130_AABXJU fickett_f_Page_056.tif
0d8ea3271089c7eed80cc4360435dd1c
41b61bd5f5c7158b8401f60244f312b56d722ccf
F20101130_AABXKJ fickett_f_Page_078.tif
2640de226816d9ebe42e240466141815
f749f7b7c3bcead6d7180a75bd71b20e97c2338c
F20101130_AABXJV fickett_f_Page_057.tif
77801a8de540643f36f6f430e81ca62a
67407ad4c21b8f30202385d274907eb102bc3eeb
F20101130_AABXKK fickett_f_Page_079.tif
620f8f3a1e1b4028e941178626aa3c06
b9aca7f1a23670c27f867da5873d8cc416ee03f2
F20101130_AABXJW fickett_f_Page_058.tif
32085eb749ae6418c6c0eafad641398a
50914d33c0d6fddc8ff08040a50e5c144b26b7a4
F20101130_AABXKL fickett_f_Page_080.tif
79467c3dbda7c4c1b90476b94a865827
039f6f8ac9e920b8e5de58dbd40ffdf75b8fd019
F20101130_AABXJX fickett_f_Page_061.tif
938c62715707e3bb0905d81c30a67fc7
1d3af7c121697d23e6c734769353582b8d4bbeec
F20101130_AABXLA fickett_f_Page_112.tif
a6385177fa25d8917ec7b67de3817873
7e5e0069d382b008d1b9ff452cbe02aec0600588
F20101130_AABXKM fickett_f_Page_081.tif
f38a5be612c30c842af7a85dd79cd799
1906338edba6e2c14cb4e626c66a99697f49a332
F20101130_AABXJY fickett_f_Page_065.tif
df8405be30c38a19c37884bfd1739405
ed91b78ec5714726da7cb2f0af5f95c0992368e5
F20101130_AABXLB fickett_f_Page_114.tif
6e0ba18afcf86f59fd2bc61444730153
67423260a1bde9f4fc203ba1b42273024d05082c
F20101130_AABXKN fickett_f_Page_084.tif
c61621b8aa57ef9f0f597ccd72cd115a
34abefb5824939387d1de51c11df7c4e517df916
F20101130_AABXJZ fickett_f_Page_066.tif
e91ea7db3523262cc49e0c2fab98bdaa
7a2a312228e3b8decc80ce381383368ee8497b88
F20101130_AABXLC fickett_f_Page_115.tif
606c1fe6675cf42c8bed836d210c07af
9610dffbb0230f4e1e8e45cb3be392d434f700e2
F20101130_AABXKO fickett_f_Page_085.tif
d878cc68adde27e7cd3e9fc07c237093
a30a20659cd9bf6a3870b406b1554e0469f14453
F20101130_AABXLD fickett_f_Page_116.tif
0e9935faf2ac55575af0b200b46ccbe6
3c1befde6f141606788c37cd89d7ccdba62b4212
F20101130_AABXKP fickett_f_Page_087.tif
39f6ace38328440158e7d3c17d638006
76b189026e2e0dc3824733202503d69fa56e1044
F20101130_AABXLE fickett_f_Page_117.tif
390f8d1466be2fb6f481df35ec650941
63df28ec255dbf95f6ece097d403582edd668ff3
F20101130_AABXKQ fickett_f_Page_090.tif
049652323fd568cc6cbc4ab9c8ed3258
89c2ac34e1bd2548994edcc301e42c9c7aeb6de5
F20101130_AABXLF fickett_f_Page_118.tif
47e27ccf4e5af0a8e68012026a84e988
dd34c953ab8c95063aed299e6651a29eb5aa096e
F20101130_AABXKR fickett_f_Page_091.tif
fb41c3176f3121e84124b304cedd51ef
bca8308768fc0c47514efc6572e4ff02a87ed80e
F20101130_AABXLG fickett_f_Page_121.tif
2d412ee136e476776f1494638cbb244c
1b0d10676b4fc76608fd45553548737b3c2a8301
F20101130_AABXKS fickett_f_Page_092.tif
66cd9a7f644a3e1030f1c922db5a3924
0f4e270aee7e196934baac26b78eadd019789003
F20101130_AABXLH fickett_f_Page_123.tif
1259e03c3fda011cc4396c4b120c13b7
c021bca9ed5e3a984846e057b9211081565b2793
F20101130_AABXKT fickett_f_Page_093.tif
e9efe28acc2022a25068a902268826bc
807438cb40386095482ab4ba4f8539d70dac28bb
F20101130_AABXLI fickett_f_Page_125.tif
f1d84c43127f7e671bd3cc008ae8b87c
f7621083506fc8b4e045f1b1ac760f19254fafe0
F20101130_AABXKU fickett_f_Page_094.tif
bbc5cb5e3e1239561de092600a346aea
f12ba216adf95df3e79c75286e86ad565be48615
F20101130_AABXLJ fickett_f_Page_127.tif
e850c835e84469b451572e5402d115d3
2381fb774cc486997ee730953bc3e4339fc9e2be
F20101130_AABXLK fickett_f_Page_128.tif
ce29feec4f49fe251548e74713acd645
e98b0592fa018c9b63f8a3dce621787fbc4e9af0
F20101130_AABXKV fickett_f_Page_095.tif
6950e6c3c98fe1183f6b9d43a124437d
9da1b7459c8c371e95b59ad778667ab7422c2348
F20101130_AABXLL fickett_f_Page_130.tif
c9f4fbc0b92f9b5d5a161587e8837658
06a0066082bdcd68a8c65b88c62b08f15258395f
F20101130_AABXKW fickett_f_Page_097.tif
24c259cd56307fccf80497ebc67a9846
a0d147cfbd3e9738c8e70a4c5829cb81e92b2f55
27193 F20101130_AABXMA fickett_f_Page_010.pro
362c09f58585cf02558a89ea80729001
d19328ed36e53e469bab24751c356827f1b73f7a
F20101130_AABXLM fickett_f_Page_131.tif
bf32c0e6a89c449adbdee7f327042c7f
72428eced4a46ec65819c53b71d6116a4e9b3668
F20101130_AABXKX fickett_f_Page_106.tif
1d545cbe4f23da1be9cfe2062d5ea127
9610a0bd57f99eb58a6a1c434d2827ded32813b4
31123 F20101130_AABXMB fickett_f_Page_011.pro
6369fd9661fad326c4cabd28f371362d
e0482a685f283906b255fc7be3cdd95bdbc6381d
F20101130_AABXLN fickett_f_Page_132.tif
add0a02037ce386f5a2f3d590f047fef
2940c366ccc56ad427c1cf8b1ca56e392a6b3d14
F20101130_AABXKY fickett_f_Page_107.tif
f182d61dded09a78c9e077450470e794
386d7feb692ae607f2e3270827b6ec2521e775bb
45188 F20101130_AABXMC fickett_f_Page_014.pro
64a3fe32c9d109edd0cff67245ee5c63
59102e2b4ef6c61468ebf2bce5c303e4c6386f86
F20101130_AABXLO fickett_f_Page_133.tif
2992cf0d566af2c147e44ab007e1f1fb
2812f7744fb5c525b85002e860bfaf4657d88114
F20101130_AABXKZ fickett_f_Page_108.tif
51b1593aeeddcbda59b6afe6e5118c8c
e61b3e2885ed9c53964570e4c0cc1eb5ab4e0e10
41078 F20101130_AABXMD fickett_f_Page_017.pro
e1c14ae661b8c20074d218f297f67bfe
40c9858014832a470a5e4d9841a3bea6a594a080
F20101130_AABXLP fickett_f_Page_134.tif
72cfbae8569b75c2d9f36f275a48839e
3c265749024e32fae52b3af20a2f7bd3e5bc140c
49019 F20101130_AABXME fickett_f_Page_020.pro
725fcb738ea6425243da12797b06af78
3d42979bea36b46c0868e2e36fd03fffb26d60e2
F20101130_AABXLQ fickett_f_Page_135.tif
5a77047a607d30d9125a298856a2628d
3e8e65037a475bff3736c4e44863f2c1fdc47c31
47790 F20101130_AABXMF fickett_f_Page_022.pro
30abdd17fe027cfa92fdb8242e958d6c
2cdeed91c840d247ed3ae7fdbc6fa8348dba3e2b
F20101130_AABXLR fickett_f_Page_136.tif
b51c6f573e170c3442e7d7831c2e8bf6
f40efebe01de5da18100f8e493baf5a0fbde2dba
51723 F20101130_AABXMG fickett_f_Page_023.pro
2968d140b2bf549ac529477ab571a2b1
81827702fef8e610f46b497e3e93543a2b9088f0
F20101130_AABXLS fickett_f_Page_137.tif
871b7c0b0e1f633e64720bd986345f8c
177ddf6d2712f682611882b93906467eec8818f0
49864 F20101130_AABXMH fickett_f_Page_024.pro
60778fc740d4bef4824df4092e3768db
e1bb41da74a7d4db3109be1cac83388d5d40b858
F20101130_AABXLT fickett_f_Page_140.tif
c1fe80350672e7fe6822d79864f5801b
e023abf754abc79118026a036446e9fdc8c7f41a
44744 F20101130_AABXMI fickett_f_Page_026.pro
fc0b974083822a19c342c3ad0068c6b2
a9cb8346d8f6ac13277eef8ae69d761d384c4d7c
F20101130_AABXLU fickett_f_Page_142.tif
522190f3ecd5d01d22d25afa11d15660
46b2b3f8ae734bb575dbefea8cef0412b2c45499
50528 F20101130_AABXMJ fickett_f_Page_028.pro
a2eedf39ea3a4728682d4cd48db7cd56
684e280a7e6e6cc74870ffcdb3c734733b822d25
F20101130_AABXLV fickett_f_Page_144.tif
f21e3e636858d388a4e0040333c50d10
be7093214e31b5f964dd1a662da584c2fd649a5a
52479 F20101130_AABXMK fickett_f_Page_030.pro
29ea0addf6b8a60860c796275f084718
cfe3342a381141ad8318f1b9897ae9fca9a233fb
52310 F20101130_AABXML fickett_f_Page_031.pro
b333e394f55d5bb26e6aff92e8664e71
5af89dccda92cf73bed720f5c528f80c1751e149
1202 F20101130_AABXLW fickett_f_Page_002.pro
da2c2dee489790f37a7bfe0e070f36da
0bb93cd681330cbffcf61f652f94041ea36a4e36
49688 F20101130_AABXMM fickett_f_Page_032.pro
3608deddd32fd0554119c6dd3ac12741
326ce614978af467775eb0b655d976ff85e19405
3683 F20101130_AABXLX fickett_f_Page_003.pro
3d536d1c1881eca4e7761ace1f1c8326
64f2d01e8c241cf996a3395ff2eaec49d69445ba
52471 F20101130_AABXNA fickett_f_Page_060.pro
b982b5443a7025430b9e8193790c5d70
07208fe58e15c4840842d0f4bda131b4deeb32d5
50019 F20101130_AABXMN fickett_f_Page_035.pro
5ca7f92f57ca6b44dcef986aec1947ad
8421bce494235c0f64ac665a54cae01d69e48714
42830 F20101130_AABXLY fickett_f_Page_004.pro
7448f15850fd5140ac9821965b16c8c2
02dd3565b570058e1d3f375ba699b0cde2070eda
50382 F20101130_AABXNB fickett_f_Page_061.pro
c8eeb664c5cc51214eeeaec60e4b08f3
e2c7666d781438009fc4a028aefb5e405b140027
26599 F20101130_AABXMO fickett_f_Page_039.pro
71b673ee7a0c2487a6b79bd00b3be628
1688b27e4f3979a01e6b3311114d001674627adf
67646 F20101130_AABXLZ fickett_f_Page_005.pro
bce5b51312b64e2eb7e8c623e7938e6c
382fc2f4d6b2c057529990405f5ed6f843f17c64
47068 F20101130_AABXNC fickett_f_Page_062.pro
4a471c43f1c1dea6696798650db63daa
09ad45519315c4fe6692190a73527ca6bad51f3f
49528 F20101130_AABXND fickett_f_Page_063.pro
b0855cc7da3a515484b0a284dc8a74ea
ed2f2632990221a79371b561e050112f3a2d785b
52814 F20101130_AABXMP fickett_f_Page_041.pro
52ea1192b2ed24ee3e0c60a07d30b586
e8f302cbd98df4c658a63a2252ad2b4894ce8fbd
48860 F20101130_AABXNE fickett_f_Page_065.pro
72e6ffbc9b66e257b012d5d5a851b86f
08b27b5cffb52833c7e593a922c3243a61d78fd8
56467 F20101130_AABXMQ fickett_f_Page_042.pro
309354be6f83cfd8d469fbcb008a9243
47352f994c7e8acf38ffe48c7e9674ac5d7d4d12
48149 F20101130_AABXNF fickett_f_Page_066.pro
e8a6138b7f3bcc1f173af61030748630
6634a24ba518fa8bbbdd001b0f0bf988ffeb2974
52125 F20101130_AABXMR fickett_f_Page_048.pro
8191846a76e361823e058b96469b7b53
5c01f37ce143885540a9d0d39b35ad23bda7a8ce
51236 F20101130_AABXNG fickett_f_Page_069.pro
1fb692a9f9cdc495caaf6d5c5964d6b9
0e236a75bf0adc43705d6bd28990bcf4d46a8fb7
53421 F20101130_AABXMS fickett_f_Page_049.pro
aec3f42d665f92ca8c2d986280aaac21
56c6c7e814c9c1ab876691a548637dbfbb6c166d
46921 F20101130_AABXNH fickett_f_Page_071.pro
ffab3d3a4ee16d9278bfec418074a0e7
6c7f50a1c5618956d35eccfa686d6fbb6f4b496a
65070 F20101130_AABXMT fickett_f_Page_050.pro
e60294f228420ab446a14f82c740e836
48ee58d602fd28ae656bb7ee22fba00aab8d81d4
6799 F20101130_AABWKG fickett_f_Page_135thm.jpg
29a4e41e8de164bb78993cc86ad7e988
41a5220c9ea32bc8ae27c74379206054b053eb35
48474 F20101130_AABXNI fickett_f_Page_072.pro
ce75f93c82c2e4369291217eab2322d4
a572e8bb2dc7912414f3089f1e94bffef5ba0b0e
59901 F20101130_AABXMU fickett_f_Page_051.pro
ec5fe602afbef0b757ba7e7f1020bbd4
02633e02c8c086a7d4e434402bb72519e487bb74
2822 F20101130_AABWKH fickett_f_Page_129thm.jpg
5701340b326b74e664348d5d1346ada9
5413ffd28b02adf26a36f4ddae604cf07001d032
50545 F20101130_AABXNJ fickett_f_Page_076.pro
05196097798c8e4fc323db74a4e0eb8b
29290f43abc702e905a69fbd1fb0078829c0c454
58925 F20101130_AABXMV fickett_f_Page_052.pro
f6a115f611c6afc34f28125e40f93a5f
bffcaeabbbac5a5ec9d7e90cadeb6bc89f8d5140
1872 F20101130_AABWKI fickett_f_Page_062.txt
6e4c9d34ed1dace72ddd8d00cd53aed0
b62dc627695eb7bfa84d25af211f7646ee1722a5
49534 F20101130_AABXNK fickett_f_Page_077.pro
456ee4e1ac0bb614408a319eb80e175d
56ddf7057dc7de25857a7170463976832a9fec7e
56303 F20101130_AABXMW fickett_f_Page_053.pro
aaf25bca9eae236bc0b319df3db5ae66
8e6908a18043a12c097df7d7b808953d95bc3113
51512 F20101130_AABWKJ fickett_f_Page_084.pro
11fa82b6e8b5fde3ae65b2714929792c
32fd133ced1c374a3ac71c220ad544642857300a
47187 F20101130_AABXNL fickett_f_Page_078.pro
0591061db0b05460aaa52c3cef3eee0f
8830209faf0d454f3b28c300792ca40d34d1fc56
15458 F20101130_AABXOA fickett_f_Page_105.pro
b4b8b04bf2eed35b9cdf10ee1af19f80
d030b82c8c26eec6f23302e1218c9d46121d8686
73496 F20101130_AABWKK fickett_f_Page_030.jpg
bf8cfe049c113fc5df806b3f96886bfa
b1061e5453f9c50650d25c758f0fad2de3035af4
48757 F20101130_AABXNM fickett_f_Page_079.pro
da25cdf3b4cbf3aec8f24684f72925a0
9fa21d3e50b6ef91628ab16d19438d75989dbe0d
45815 F20101130_AABXMX fickett_f_Page_054.pro
3e16fb275f298a622f951917f7c184bb
560afa6b6801dcdf58cd7ca003870cc7527a6cbc
23469 F20101130_AABXOB fickett_f_Page_106.pro
6151b4a0ffd6f6c2c919819591fcb488
f16b297721efd77a55ddd11dc25e84975441afde
F20101130_AABWKL fickett_f_Page_111.tif
36d3ea9d667a3bc9009fcc4b51a1b75b
de5f5269048ce238f3f5880519bd6f716b93a274
49809 F20101130_AABXNN fickett_f_Page_080.pro
a5f646c1ca4cdb0959326e4f3c210dd4
d916b30022125acca90ffbd70b537ffb0d3d970e
30008 F20101130_AABXMY fickett_f_Page_057.pro
b3212831820b85523a5a091beb3b0510
d0ff5e7ab3c44d791fffac4045d6a501a3fccd71
105959 F20101130_AABWLA fickett_f_Page_015.jp2
fdba8e2ff5d5573340d8560aec890447
0248be3a81f2eed5370b1f58a94344b45de2e289
24367 F20101130_AABXOC fickett_f_Page_110.pro
8413e1ac1e33e3d6b6b6f413bb773112
c5b293647916afbe299f5e1db11a98c273590b44
F20101130_AABWKM fickett_f_Page_109.tif
dea04dd1efb979a109e8f3b503a88acc
5988d12c763fe3b7852146e867a6eb4c1f19e84d
52167 F20101130_AABXNO fickett_f_Page_081.pro
5259458bad1ddf94191b12f8e86f14a5
2d7823760fc81db5cc79c8f2056d638c2c1d8424
42558 F20101130_AABXMZ fickett_f_Page_058.pro
050d9bbd9294810f184bce1bb026c4c0
1c1a3d116c4b43a0df2dd353c7664b14446711ff
6447 F20101130_AABWLB fickett_f_Page_036thm.jpg
93677a0e9747c074ed44f8fd51eb4caf
89799953d80c5fc4b5035b542b235ce3105ffc33
24510 F20101130_AABXOD fickett_f_Page_112.pro
399bdbef5b8ce8f32150640ffc0c49f1
0650b08d82cec316b4860a525ca42492fae02cdd
23679 F20101130_AABWKN fickett_f_Page_061.QC.jpg
d4dd25f5b69772df21fd6b14a7d8676d
19ebf8bf21692ddf4f90de833677e137a4eba42e
48181 F20101130_AABXNP fickett_f_Page_083.pro
bd7a990fcbc01a30bcd12c02af44a00a
34e649806f78c4e70d3be960d7e158af71b30cbd
24268 F20101130_AABWLC fickett_f_Page_075.QC.jpg
a1fa44b2df779b94cef296fc4af3a23f
b90a9c8d66474ee84eabfd7dc336eaf5081371d4
23655 F20101130_AABXOE fickett_f_Page_113.pro
fbcb7e4063ca1998df59533daee7a01b
020a064ca393b01b74a2155d2121dba383064d2b
61271 F20101130_AABWKO fickett_f_Page_099.jp2
76cd568bd2437a55a100cdc0265e68f5
87c86275acd69eb6b2c862bed5af5a29df89e8b3
48882 F20101130_AABXNQ fickett_f_Page_088.pro
f8c63aa715a86f6a1e21c80bff79f538
5faf974a6cf7f4dbd0eacdb25340f46d460b88b0
111821 F20101130_AABWLD fickett_f_Page_084.jp2
1e36d79ba39537af62fcd49462056bf5
f359977cb180d82ee5e40e0a45feb0937e8c0905
25809 F20101130_AABXOF fickett_f_Page_114.pro
2969d2a2a390f726f1e28d92f2513366
47ed59318fdbcbb20571783f3df5c92bfcd6744d
69808 F20101130_AABWKP fickett_f_Page_004.jpg
aef2f2ecee9f44e46a7f83161e219393
de6db2026faea48ee6b7ff08e3ad2880931d3a53
51265 F20101130_AABXNR fickett_f_Page_089.pro
352a7553caf6c1d822ff747017f9ef3d
3a0f423b21a4f62a66a0b986fb4919d5df699373
22469 F20101130_AABWLE fickett_f_Page_078.QC.jpg
eaab63acb4e1826963538cbaab719882
d29d8c4bc2c3a3a8698d26816cce4804693702a4
12844 F20101130_AABXOG fickett_f_Page_118.pro
8fc1bbd1e22f7afd00eb6d5dd7d55092
410bb8f7fbb1115bf89cd9e0561e2f4747f3752b
34556 F20101130_AABWKQ fickett_f_Page_099.pro
4cc34949e3a14f819d7a91e59aa5e7d6
1a0b5db7fe784372ea3561767a1da977869d9b0a
50068 F20101130_AABXNS fickett_f_Page_090.pro
eef54565c8b6107c0f54947a25f4c982
1642522a29a7d112fde67d9e7177d40b3b3b0d17
50132 F20101130_AABWLF fickett_f_Page_033.pro
7f85c531d7e10885f2df49903e25ec5e
12476a19a438dd0ff677b5c51fdb46c66eaa4cf5
13360 F20101130_AABXOH fickett_f_Page_119.pro
105f79d2099f88136eb37689a984670e
a447cfd2f688665f1a8f4d5edd7eedd11fdee143
1051954 F20101130_AABWKR fickett_f_Page_005.jp2
6fc8e4df9021386e5ef4d98a9b25f8ee
e5fdf0a105c206815b3f005dd026ff37c0fd4184
38918 F20101130_AABXNT fickett_f_Page_091.pro
187f93244e0990fcabb9b5611564ba6e
08263a0f6cf2c634165d164ce433c92e6269cfe1
2413 F20101130_AABWLG fickett_f_Page_136.txt
9affbb17aa9260395aaacb210c99957c
b6fb991ea11442bb454121edab711ff753cabb2e
27014 F20101130_AABXOI fickett_f_Page_120.pro
6edd07f65736f63aa41a1b26ec44c694
c6d2603532adcca9840b7145c769402dc2150c1e
54257 F20101130_AABWKS fickett_f_Page_044.pro
cf5292b85d71a63fe8606a71d97c303d
10bc0af68c8b2b8a14bab681f49dfa073344b487
45354 F20101130_AABXNU fickett_f_Page_093.pro
1b885cb7764fd84692ecd36cb93f924f
fbc78f0c55589d297464f2afca95f1faca6b79a2
1447 F20101130_AABWLH fickett_f_Page_128.pro
cb9dee966c686db3c0ad17fee2bcac4f
58bf64463b23be4556d1bd7092a699db8827afb5
18317 F20101130_AABXOJ fickett_f_Page_121.pro
516c20b8d33bad34f5e09a7665d5c2d8
ca93d56c79698999d9d65540745f97bf09b8df08
2076 F20101130_AABWKT fickett_f_Page_030.txt
e0a31988eb61401af0643711c6052cd2
b846f7d66f73bedae63fdfbe9fe20674e8b41fda
52291 F20101130_AABXNV fickett_f_Page_094.pro
e26a8c7c680f6210c945f77a56503dcf
8e79e70f3aafb5dbf56dea4257db102845a33f1f
7228 F20101130_AABWLI fickett_f_Page_130.QC.jpg
6c1a1c4ead7904ff9603e693b0a2d706
a92a44d12d3dc88c3159f52ef81330d03ba7b560
11651 F20101130_AABXOK fickett_f_Page_122.pro
edb3fd8d2f1ce3537a3e6699794ae21b
fe620331c1af02309712f5fff2c27cd84247c3f6
24112 F20101130_AABWKU fickett_f_Page_025.QC.jpg
afa9f1c20a27339a8e06f7fb13b3db27
73b7f604a760b40f78b2b305dff9ec0eb68579e0
36751 F20101130_AABXNW fickett_f_Page_096.pro
1d51cb5bb99b541aadcd16b672893074
8d5d1afe06eb673542cb01bf82d33f48cf5c5354
30671 F20101130_AABWLJ fickett_f_Page_103.jpg
17323c1bd7e1afa4f6a80ff95ed0a3cc
638835178e4098d61205ef60fd5a010d8216d7b0
11807 F20101130_AABXOL fickett_f_Page_123.pro
0039ae3a8fc4b097e427dbb40b961891
1dc6a535e5a01bf2879a0f1c5c7377a47b7c1c57
5801 F20101130_AABWKV fickett_f_Page_009thm.jpg
2d2c0578a9241fb34d7b60bb959e0207
b8d8a2a6a28e429a0434037e485128d01cf712f7
47471 F20101130_AABXNX fickett_f_Page_097.pro
9cf40a747d6f506d638744ab2e42080d
fd47357b3b4428c9151e6d8c36b83600a5ec6b8f
1731 F20101130_AABXPA fickett_f_Page_004.txt
7516fd2f0ef73e872783ea4e836e68a8
2aae538a25319ba49626aca0d5e0251a3feabd43
11697 F20101130_AABXOM fickett_f_Page_124.pro
4841187b72fbc69aa4578faace0177b9
77c5d44432cc5143c992db476fd86cfe41d60500
48677 F20101130_AABWLK fickett_f_Page_015.pro
32c5bb9c2710b3fa4ff98ee9522b3390
788e6e474745e18f45baf907a0a6d1bff0a4cceb
2781 F20101130_AABXPB fickett_f_Page_005.txt
f0e0c11411b6e318673af6869a0da583
7ab84f759b5ed826084d30e9bf43530253f9605a
11396 F20101130_AABXON fickett_f_Page_125.pro
de39728636bb4580fab8644e548f5370
2e3f34c5ba982a75b74dcfb0947de8b4ee7554dc
33520 F20101130_AABWLL fickett_f_Page_131.pro
bddc92196b037bfd50fd3afb1d76d5c5
ac97f5add753c5edb01d20ea44ab4f35639fcdc8
40271 F20101130_AABWKW fickett_f_Page_093.jpg
b220fd0c55ba0ea33ded551b16b99903
d2c568153709516061017bfe3d3651471bfdeaa9
32605 F20101130_AABXNY fickett_f_Page_098.pro
0e96ac69ac9130d34fbd3002980889fe
a1f680aaf0db1458a64479db33780cec360d4d7c
2481 F20101130_AABXPC fickett_f_Page_009.txt
01bae397c5fe98f6fab892fd34b79fd8
30ac22a2e43736cd1675d3ad2995c0ddf4a592f2
31546 F20101130_AABXOO fickett_f_Page_130.pro
70edde88ebf3b11fbe352b83d9ffa2e6
bbd61562c18926d1f1fb7d483d996b8e37cdc215
17262 F20101130_AABWLM fickett_f_Page_102.QC.jpg
cece1ff1976ff6dd846c58a72298bf3b
1f8f37caf9bec542e07fff7757bb8f4afd311798
F20101130_AABWKX fickett_f_Page_066thm.jpg
c24611e942a15c8596e301dbbbf9597b
a0b71750f04a5ba7cbad9b1e22c20116372f7862
13206 F20101130_AABXNZ fickett_f_Page_103.pro
b3f999ff25e46358e4356a1f9aacc7fe
ac85aa5b5c7bc1d2572024c0d88a80e0ba237639
71116 F20101130_AABWMA fickett_f_Page_065.jpg
a34b05e39ceef18d59abb4fe7752067d
1ebcc46d98b2358ddb60cd967bea10d34d476eb3
1977 F20101130_AABXPD fickett_f_Page_013.txt
d075dee42d2864bad8300f7bd7c8ddaa
d9c7b77ceae8ce9dbae103c45cb1a744a3819c2e
53829 F20101130_AABXOP fickett_f_Page_134.pro
f5ef7b12538f470339f82b2ae0f8b816
baf78f520e52959fc289b05747b20658d62c4978
63720 F20101130_AABWLN fickett_f_Page_040.pro
72fbbb23f26a2979d39957b26bc33127
050954eaed3ea99ac8385a0fe8f23359bb3a75dc
72062 F20101130_AABWKY fickett_f_Page_068.jpg
2a7ef13d94a35faa7ac2a38ced66300a
1c524577a0aaa0b8cb2df7dc69d136e84b930613
105774 F20101130_AABWMB fickett_f_Page_067.jp2
33282b4f84d56dab3427f054e499b83f
a99a87d4648311b4b4014f2921c5eb8c9d9db6df
1854 F20101130_AABXPE fickett_f_Page_014.txt
6babc48e0c246d85dcc2b5f4b2bd35fc
90d4337bc0340dc87ad30fe513d7e33d34e84525
63433 F20101130_AABXOQ fickett_f_Page_137.pro
a2a973aae51b099845e2a719a473bacb
771fb604328f321bd706bf3efee3d3286ac4b4cc
16089 F20101130_AABWLO fickett_f_Page_126.pro
8fb87a57539cde9597e63b5577f7d0ea
b191a397e3d51787b75e57d1aec8e2980281cb62
9353 F20101130_AABWKZ fickett_f_Page_052.QC.jpg
bafa60561e421fe841bd52d5bddbb989
d47018ec2eb7af8bfad52b96d223ca18688bd9da
8410 F20101130_AABWMC fickett_f_Page_055.QC.jpg
c64b961d1a7d898747ae01cee5876a07
5c7f5ce0629a87a1dfe12358d82b30e0ba5d4e2e
1918 F20101130_AABXPF fickett_f_Page_015.txt
66f94d08ad7e93e1dcca98131a1414cd
f4e2e989bffc2d599f6d7c2822e8002d95406565
66338 F20101130_AABXOR fickett_f_Page_138.pro
2dbb0c633521bfdde5a29ea2716e3ad0
2363ae186826cc4f1c6fb119b5323f67c12065f3
111216 F20101130_AABWLP fickett_f_Page_023.jp2
f1666c60bdfaa6493a63547566c6e946
8a882a9abd5325d7bb1cabf614e3e1035b95cd6e
71925 F20101130_AABWMD fickett_f_Page_077.jpg
453d1f908705a715dbb44834f54aaf08
83f5eb647ab54c5f0fa26b3d9183a872ff509723
1728 F20101130_AABXPG fickett_f_Page_017.txt
dbe75ef953667ba5d807e53d6c0ffee3
4418173c58105fd0e6bc61070b526d4d274d7c90
64742 F20101130_AABXOS fickett_f_Page_139.pro
0a14c0cbeaf07255e1fef42c370c5986
cbe8149f4b02dadf5d4ac69186fecbbb12174686
50289 F20101130_AABWLQ fickett_f_Page_025.pro
c3e2cc74ad615e5c2a528c8c8f1d5dc8
880e33c7d38c1ada697942190be7438d5dd30b3f
401347 F20101130_AABWME fickett_f_Page_122.jp2
2da669a626dc51b80086e468d1c6901a
15b96c592caf128d23a8daaf426771602fece478
1997 F20101130_AABXPH fickett_f_Page_019.txt
f2e0615f314f16d4cd29ca4cde6f7107
104ca06a888aa6e830b2193c1982dc8e17236eb1
69001 F20101130_AABXOT fickett_f_Page_140.pro
bd9d4457c1c31eba28dd6f61369cf90e
8f3011762d7c06fcbccf0cbee2637074529e9701
4259 F20101130_AABWLR fickett_f_Page_008thm.jpg
d17e76b68810a17e858c4c9b0887c151
f2fe0d29b9b1b02886817f389891b35d05ed750d
24694 F20101130_AABWMF fickett_f_Page_130.jpg
8730a6576327b42e694e2e06bfb142b7
b65d6ebfd28baf1282988a5085d868398695c8bd
1924 F20101130_AABXPI fickett_f_Page_020.txt
bec6583f8849a77e6fd7e3d573c3581f
c679877612effedb7e02aba650e4b8f724c294bb
65544 F20101130_AABXOU fickett_f_Page_142.pro
bc2649de0ae1f2f61fa7d6d96d667c61
d5dd138b5ec9aae8004ac30c8df6098f83a359aa
F20101130_AABWLS fickett_f_Page_122.tif
4d201c3010f904c477c37c13cf974620
23f657b1f23498abb2c5723575dee0121daae8c5
3260 F20101130_AABWMG fickett_f_Page_126thm.jpg
3c1129d2a9369f1411fbeac87357d3f2
efe75d28a6e0cc7ecc1834e6509944364bd723ca
2055 F20101130_AABXPJ fickett_f_Page_023.txt
d196faf675a31315738a8a351bab853f
7f983d7b58feb2e8b909893fc49f55abea07dc16
66349 F20101130_AABXOV fickett_f_Page_144.pro
460da3a7d34c1673075219f8e091007e
3ba7a4c5f3582609d70d19f486131b6812c6cab9
74768 F20101130_AABWLT fickett_f_Page_084.jpg
a656ea8339f0529ed16eaa25cdef280f
e199b2cca03a09dac6afa2fdf684459c29253450
90255 F20101130_AABWMH fickett_f_Page_040.jp2
ab5c1665826c81723988c958c4d035ed
8434188d5cecccf77e75b909939f5c426bb11d4c
1970 F20101130_AABXPK fickett_f_Page_024.txt
7037a7ae8a4657a064b6b613c7c52a10
195312fcf8d35d1bd45764e1fd79fd782f50eea9
6943 F20101130_AABXOW fickett_f_Page_145.pro
6e4ed4ee2d324a81c2b184877dc5b8ae
82b2adbab188b8ed1bec6f81d4d2eddf94443442
70049 F20101130_AABWLU fickett_f_Page_047.jp2
70b33965a225f0f3043d169d6665f7fb
6db81ac3a9639eb59cfab4c4f30d56eaaa482c61
71984 F20101130_AABWMI fickett_f_Page_079.jpg
0b889e8a2b5f94c4fb2ce247505d75b6
5ba11a8c0be96df34be3482c2b890c5d9e9ddcc3
1989 F20101130_AABXPL fickett_f_Page_025.txt
659d6fccbce9cb76a58c16059cff23ae
28d8c25453afa80bd5bc3c8fa9c8d45636d9ad9b
13534 F20101130_AABXOX fickett_f_Page_146.pro
bcc2d3c65b2fcf56bcdcf7164c3d4288
6a1de9309c3a17aabc6130dd683f8db7f0b42d9e
51305 F20101130_AABWLV fickett_f_Page_073.pro
bf62549e52c847613f584e6bd4089301
b53a0b9ac57d9b2ac1bae3fadeba1fd2e36ef066
4151 F20101130_AABWMJ fickett_f_Page_039thm.jpg
a1ba3952a5aad16c52141e4bab15827f
805acdda9e6284ac029976dcfe02285481db8b10
2772 F20101130_AABXQA fickett_f_Page_053.txt
aea92c9d0faff130cd898849696c770d
9e59a8ae1cc86dddfc555887ec907b32c3b129f3
1987 F20101130_AABXPM fickett_f_Page_027.txt
a62516ab1e9d25b048dffe053675ff47
4c78faab35ef4537e01f1bc7f6665a47d8f217c7
524 F20101130_AABXOY fickett_f_Page_001.txt
cb7244126cd21c3f086cd9cbb2282635
2346e34de71439a16d31fa5f074eff8626a84031
63858 F20101130_AABWLW fickett_f_Page_135.pro
5868fb9b8b737550e3c2530e0e3be726
14637a950d8ab62817ebc18c662e492f90611746
49136 F20101130_AABWMK fickett_f_Page_059.pro
658d9619bfc0a20677affdff9b735b31
8df45fad57475a7dd6485d9318e04b47c40d9cf4
1756 F20101130_AABXQB fickett_f_Page_056.txt
c19379a3df6a120cdc3413efb7a4c1ae
f9b2febd30c62d4850cf750cf61f42504e1e1bf2
1808 F20101130_AABXPN fickett_f_Page_029.txt
39810c0cf6f86cd3240bdcfb7807deca
571b19d63f43b611e82fdeb998aa46cf032c8ad4
1076 F20101130_AABWML fickett_f_Page_010.txt
8a520d48b6a9d057c7dc156054b08719
b384ac9471ec702da6106923eee9ac8137117c0c
1483 F20101130_AABXQC fickett_f_Page_057.txt
695a250b8a82b6af5b9ab078ba654339
5be5e7132b9ef055ef728dae44602a2b5ddc4d5c
1959 F20101130_AABXPO fickett_f_Page_032.txt
6c120cdd903cf8659707932ff0c4c7e2
0c2f52b774dfad76ea98392eaf6724a32bde4187
111 F20101130_AABXOZ fickett_f_Page_002.txt
08c98f4465e0e047a71b4af8d77624ae
70f3e627dfb37d489685478451e67ac6da1909ac
560262 F20101130_AABWLX fickett_f.pdf
b660fe7f9a582d0df39eb2e7458608e8
4e73d7d14f312a6d6a28aebcd8a82e7512283845
F20101130_AABWNA fickett_f_Page_062.tif
5a7f2e494d55f4a0a7a25d9dabc3958e
2f93f943c393f52cf1b12f4035eeef6557224bfb
F20101130_AABWMM fickett_f_Page_100.tif
6f173f392d2ca0b859868561bcb4917e
843300889acdce61fd433165f29e7a5f261bb5a6
1784 F20101130_AABXQD fickett_f_Page_058.txt
cc023d9fd0fadc053c08a394b3175737
4596b0be313287650530a3a303d0da6c1c1b8dd6
1974 F20101130_AABXPP fickett_f_Page_033.txt
bb0cea53e933a3398abfa1519497e8be
82dfda05250cb878894d9fb2ccfdb46c4b127419
9205 F20101130_AABWLY fickett_f_Page_124.QC.jpg
e5587e158fa3885a911a9456020c26c7
f0c373dee128c4f8c3d3615897c4f85cb3083c64
59354 F20101130_AABWNB fickett_f_Page_039.jp2
cd36f4f5f9f831fc05c2f3d567a7f281
766edf0d6d93ac38f47221e3493c456ef13669b9
6694 F20101130_AABWMN fickett_f_Page_013thm.jpg
deabd7e1efaa081dfe054e968ca8114b
9d0324bd5d64cea5445c8b677165e9e13311b27e
F20101130_AABXQE fickett_f_Page_061.txt
5d7cc253a7afcae27818fa4b6209fb4a
365d0820b965b4f719e82bd79ce68911718bda57
F20101130_AABXPQ fickett_f_Page_034.txt
46c84679e87a4f29ee229591ea665f61
a6f5034e66ced7bc79fb0dac1e77d8a690b5d495
23106 F20101130_AABWLZ fickett_f_Page_062.QC.jpg
e82c73a2682c25893396249ace19bc9a
2349677576fb766350e726b18f14289cdd541e25
50471 F20101130_AABWNC fickett_f_Page_027.pro
afc235b41baa951efe29fef972b4fd7b
4c7afe23d09ded1c3757b7b3df0f47e039840699
25831 F20101130_AABWMO fickett_f_Page_133.jpg
8de100897e350629652aae43f884cd85
341ed956b3d0095d2eb8ba0d5b4106fb4f2dcdc0
1957 F20101130_AABXQF fickett_f_Page_063.txt
58885c44d14b27df17e9df437202c32a
5c0442c34a244b2e144e18d296d8fbd68f2166a5
1963 F20101130_AABXPR fickett_f_Page_035.txt
ffb69d1885aa5199da0b071da2c19df0
45725bdd32166d5dbf0d7e15d9f85a26c70d13fa
24359 F20101130_AABWND fickett_f_Page_084.QC.jpg
90ee8b5aa2f8141bbad8d0169bd680c7
1f75953ba77c80e5dd25d38a0a503e19d00ac2a6
54648 F20101130_AABWMP fickett_f_Page_113.jpg
2484b79f46644c9f765589cb32871c70
4bb3260070faef1a0b1df7b66ab496710b8164a5
1899 F20101130_AABXQG fickett_f_Page_066.txt
517efeead79912f7818daa0a2ae59a5f
d3098af62964ca8c76f3d20710f809cbb71c0f14
1976 F20101130_AABXPS fickett_f_Page_037.txt
e6392c12eaf48b5967ce322f86cb4a23
e4fec85acc12ba1ae296b3668bd411759bb9b4e6
6590 F20101130_AABWNE fickett_f_Page_062thm.jpg
63f03d1c348e1190bf82df86994d2cca
54aff4a12800f1f0203c695c374a23b602c8f396
F20101130_AABWMQ fickett_f_Page_060.tif
c78b3f04209a00bf2752abf608808ee6
2a62ffcdbc9b2aea5885dc325c41b78b61270dc1
1909 F20101130_AABXQH fickett_f_Page_067.txt
c2d098459fa27ae8e37284b92d8a79c7
65c72f649ed418a2073295ad8264232fcfb6d606
1922 F20101130_AABXPT fickett_f_Page_038.txt
6fb28a544f8818528e400f8ba3951445
377f6aabdeb00551db94d925ff314169a6ac5216
23599 F20101130_AABWNF fickett_f_Page_065.QC.jpg
7ff4eb70f1379577d6fdb5f1c2622894
a65270807aa5d8b0329a8e4f25b75abee1ce3124
2689 F20101130_AABWMR fickett_f_Page_047thm.jpg
bef327f050b864805e2deb9d4e73c214
f09f2f00b19eb3fc0f48c82c496bc2b33061fa10
1852 F20101130_AABXQI fickett_f_Page_071.txt
b8b7d90c576fd8c53c3121be06a73b37
29b6f325d2687896cca0e42bb93d86c07efe9e91
2621 F20101130_AABXPU fickett_f_Page_041.txt
95e6ef9f7a6c442216520ea749f8e10b
8a0d8e200ac3533d30afff95e0b9962e9c8fe19e
105436 F20101130_AABWNG fickett_f_Page_065.jp2
a75eecb2c6da181a66f049abaedc2412
4151d9bab85483a799125d1ce0963e2a9d2f7bb7
11100 F20101130_AABWMS fickett_f_Page_121.QC.jpg
87ac1f9285977cee17610b178631c983
ec603c57882ac67ef0030e47ba95a29e839abd2a
1908 F20101130_AABXQJ fickett_f_Page_072.txt
d1b6ef65a97bfb65ddafb1419728264b
3d72ae88e2e0e7decf22146dded0673a2d295b0f
2768 F20101130_AABXPV fickett_f_Page_044.txt
203d6f364fca55f773e2b6f925fad469
d3313440e38e12ae1023c2b687750a7b7169e4df
3616 F20101130_AABWNH fickett_f_Page_093thm.jpg
236cece1aac135daf5c0b9c906219487
66188431efe34c925829418227d36a165dc6d27f
128781 F20101130_AABWMT fickett_f_Page_135.jp2
04a7f4e18314f642ef855793b350cbee
ceded6cf84e547cde3f004b7c6b892d9dbb547d2
1993 F20101130_AABXQK fickett_f_Page_074.txt
8c47a7f7883b3acba0614b2a2d9dbb3c
37a86a5a7f90a6024402bba2ecf86ace5f834fa1
2778 F20101130_AABXPW fickett_f_Page_045.txt
4a0972b0343e1691d3b1774db2f605a3
3ef48e23865779e756f934751bf130677e912a04
F20101130_AABWNI fickett_f_Page_138.tif
8c0dfdd13fcf0e426692596440f0f7ae
437f3f5901bb7bc2f2bea19ec3c985a0e3e835af
2992 F20101130_AABWMU fickett_f_Page_052thm.jpg
0c7963fe0d35822ddaa377e618b16d69
d3360ac0270f69dba39389d30a0de98c19ed8b0b
1945 F20101130_AABXQL fickett_f_Page_077.txt
3ce981c1003bff79caa6385374a96834
47accef187c4ccdc985b51ab94ec0c4a3cb67d60
2404 F20101130_AABXPX fickett_f_Page_047.txt
8ac65e34180fc98eb2f2d7f2fcdd2f62
8336fde0d6eea318fd8410d703f7d1b0d85e43d0
F20101130_AABWNJ fickett_f_Page_012.tif
46026668f722311889118f77cfeb2935
4f836e012998adc652a53cbcc1111ee0d57c19fa
2199 F20101130_AABWMV fickett_f_Page_134.txt
e1896673ac809d4b9298e478b01e0437
39496df02a5db3eeb22592890f9cee227e4fd3c6
964 F20101130_AABXRA fickett_f_Page_102.txt
1556175a1c85284b1b7338190c05b847
05e4dc78e0233abc266776e090c56507bd7b1455
1956 F20101130_AABXQM fickett_f_Page_080.txt
0b3e074508068df9209e4af656c4d022
35cd913e28063d982d9781158df1b7ef09c85a89
2770 F20101130_AABXPY fickett_f_Page_049.txt
737471cc3f7ba8e9ac63d570b411e11c
0062b2a446a809a5a5a7d0852912c6ebcfabadab
1527 F20101130_AABWNK fickett_f_Page_006.txt
2fed717793ae73c082cf9eb563627033
a125073a4d4dbebcc114048925f5c8885f70fb31
3504 F20101130_AABWMW fickett_f_Page_108thm.jpg
06e06926ebc17af68b734ff9801730da
cb227aef3b15ad9d5aac71b3d2b88f92905ff55a
F20101130_AABXRB fickett_f_Page_109.txt
48931e77ac29ac52e051a5c3057fff9c
94ab0604457d3fb7c38a247c155c4d0344c633c6
2004 F20101130_AABXQN fickett_f_Page_082.txt
56f5124f7ae5d618a9989515a6b49983
3351e9b972342f6cc4c1c2b48721777459434200
3383 F20101130_AABXPZ fickett_f_Page_050.txt
44aab6d2650e4bfacfd909faa5e3d74f
650cfe355770113e36716e6d684b87bbba182a38
6475 F20101130_AABWNL fickett_f_Page_071thm.jpg
5396c18163c28aa50bcecf9098223f5a
4528e303ebd6deefe1949f4c81d8c643e72a4539
1935 F20101130_AABWMX fickett_f_Page_059.txt
b0f67fea7a4b0f6c40956311dcfbefbb
782dd326597a9da12d18161de7edb479984cf90b
1505 F20101130_AABXRC fickett_f_Page_114.txt
ce56ef18a306301a52d8aa3e7b45a371
85e893220c164ff8afcfe98cce996d8b05aabde7
1895 F20101130_AABXQO fickett_f_Page_083.txt
3b880f1f424cd474faf51ab003143b06
e3f1bdcd7954c109c681cd9a1877ad077ffabbaf
F20101130_AABWOA fickett_f_Page_054.tif
8fe71d5aedaf1f7838f18fb528ff8f8d
c06452aab62c81ddab6459384a9f20d7211035b3
109371 F20101130_AABWNM fickett_f_Page_059.jp2
dda748f0d1a0ff21541d297dd2e2bbbd
28abd1af3a2eda3c0d5026883ece52ba6bc1e1e1
775 F20101130_AABXRD fickett_f_Page_115.txt
89b913688a48ac3d54442e1e5e01a077
1ecfb08a40c0f9d59abeb668aa19e99f98738153
2021 F20101130_AABXQP fickett_f_Page_084.txt
6cd31399264020bad8bdc04c50e90492
fc8c98bd93e097f1888b7ce22fde8b8a0a2dbe37
25678 F20101130_AABWNN fickett_f_Page_109.pro
b4e38ad425e00acb8574f329069e3b2a
83122e8baccbeca7869402ef4b8e46069d43add2
6822 F20101130_AABWMY fickett_f_Page_023thm.jpg
56eafc0fdc392560f707509a30821145
553dfd6bc5253447fd7c52ec64627dad3994181f
F20101130_AABWOB fickett_f_Page_104.tif
37233ca9450b5ae80efa117690112332
7a7ed82a4d0e972c63625ec66b05078abeaa7a58
1132 F20101130_AABXRE fickett_f_Page_116.txt
f18bc57c6d6bcf03fed2154189a9b371
86b712fbacd118721141ca9b833b90c4aa41d4fa
1938 F20101130_AABXQQ fickett_f_Page_088.txt
78fac77903b108d4cf8ca13b3e1899ee
2d46af88f3c1a2a4fa7033504498e1689407f6f4
48633 F20101130_AABWNO fickett_f_Page_038.pro
f9a6b3ec3810d22c2ff843b1404d1cfe
41141642b00820dcc7736cca982cf1a9c530b1b3
98735 F20101130_AABWMZ fickett_f_Page_026.jp2
3bb46e89ee00cd07c0435e20c476666a
256bfb38d37e6bb74ccc8d7e8031b401536d71b6
65604 F20101130_AABWOC fickett_f_Page_143.pro
c35ee2e8fd3d75cc4d07c3ebd7f29447
c5b9eb952462da8796d5ff504b87988712cda7b3
725 F20101130_AABXRF fickett_f_Page_117.txt
e8d3128cd79715e7d1fb208327cd873e
6e18840dfa1678394a8862401d10f2fbba43f67b
2016 F20101130_AABXQR fickett_f_Page_089.txt
0941f1fadd60b1cbc94dc31aed373f3c
8066e74229c6ed8e32d3b081347ca0d4cc66cb97
111826 F20101130_AABWNP fickett_f_Page_077.jp2
0d9d033297ae5afc50a0de37eb890ab2
37b16164dac58dd72c094dd8427bb73348e84640
6469 F20101130_AABWOD fickett_f_Page_037thm.jpg
1b1461d87c12869c2516b6466e89ac72
133c47887c8899c64d3614d2814b707ba27e8cd9
1411 F20101130_AABXRG fickett_f_Page_120.txt
d3f6226839934ef528642a93072582b9
13a09a6188aab0c7558699a8605a25b7031fd279
1962 F20101130_AABXQS fickett_f_Page_090.txt
575709ade92b5f7706d81207108078ac
528edc9a879c3d93d3090d63262abc0d5399b4b6
2581 F20101130_AABWNQ fickett_f_Page_007.txt
16f61a1d8fd1090a07d92ff95d610c5d
d1d4908d07d438b92fc52d036fdcd9692a88d79d
38461 F20101130_AABWOE fickett_f_Page_008.pro
a24678049bf855f105fa2a91967205c2
33053a169ae3fe67294a2adf622928c4005d4308
989 F20101130_AABXRH fickett_f_Page_121.txt
21e5f3ff09e88f4059c78a61e3f21fb9
6c9891b550e0621ae3f9c1500fd403514f194f08
1855 F20101130_AABXQT fickett_f_Page_091.txt
5a12653b6be6edddc61928d555912004
76ee676897d128b56a3e7c1fda057de403ecca9d
1778 F20101130_AABWNR fickett_f_Page_026.txt
de2fd8fa9da25302d7152a8b91b5c629
287b3748322f3d64b2fbca67c58cebb8f01eb5d5
2637 F20101130_AABWOF fickett_f_Page_139.txt
709a8148e2bebff602ca8ff7a97617fd
0ed8409ff69c8b80c51c3dff8fc3b88ff67ebcbf
666 F20101130_AABXRI fickett_f_Page_124.txt
ed65d2fe8a9b73e26087eec433b94ec9
a21810dd2499a229db05ced1ba27117fce47db81
2009 F20101130_AABXQU fickett_f_Page_092.txt
b601510b9e9ffee7b8c18d312a26188d
fc5e3724e853f9243864c1ab5940bd7f448b9c31
22169 F20101130_AABWNS fickett_f_Page_022.QC.jpg
4ac278c2f2713867d3d84f9342de4e35
7034f36e4e1376e48b68b27a3d196d49b494eae6
1896 F20101130_AABWOG fickett_f_Page_022.txt
391e39b2e89f3662c656d8bc82621f8a
ae6450e3fec64727383da174dfd43f62b4d814bd
838 F20101130_AABXRJ fickett_f_Page_126.txt
2838882b9f1af750ad6de170fadc0cdf
61dd146b6554c2322bae02a087dce67e50202bea
2257 F20101130_AABXQV fickett_f_Page_093.txt
eae1431ec53b9803710d7ff3573a3e84
bcd7dd573d378dbf4e0b692f918a566ec14f2583
66073 F20101130_AABWNT fickett_f_Page_141.pro
7e22c1d28ffb047ac1706800ada33816
c74c1273fc045b5a5cd665d0979720eb82328752
6723 F20101130_AABWOH fickett_f_Page_089thm.jpg
11f778258cd2bb5b8575ed49f0a98a10
eeedf811aff05e962c64f2c1a5a2f7efbc73d111
1687 F20101130_AABXRK fickett_f_Page_129.txt
7a9b64fdddaf420e4810939e344c787c
11a4d746472680865f0207d38b8ed8bf749e56b2
2440 F20101130_AABXQW fickett_f_Page_094.txt
5ba5d7e99d0893b273b8e2053adc1a8a
0913a6e445998f25b23c8765ff95b872358769ec
F20101130_AABWNU fickett_f_Page_124.tif
7d4181672802a7fd9b5cb26032e44173
70e529eebda0429df3a1600a39562eb8e9768f32
51504 F20101130_AABWOI fickett_f_Page_086.pro
c892a364e0513451654a2d6930bcd18f
4f997842fe97f9a60af070aecb09e6eb383ce6c8
1552 F20101130_AABXRL fickett_f_Page_130.txt
cafe70bfa0795e3ea4c88e79973a6052
02b1b35f6f471aa76eece005c6a830e8ebf3f8a5
2709 F20101130_AABXQX fickett_f_Page_095.txt
1a50d24e360f0183511007d5f67f1d7a
ab1085ad93a8c2d0f4febf7aeffdaa65e482643b
431922 F20101130_AABWNV fickett_f_Page_121.jp2
0988c2b24324273fc787dbaecc939de7
c359e520d938c227632fd3fd627248eb6d7f6711
82895 F20101130_AABWOJ fickett_f_Page_046.jp2
8ed7f3e430dc362bdb5b55713b7f39eb
69646ca0ae4d1e9f85d9bb5792c8e405e19931e6
12086 F20101130_AABXSA fickett_f_Page_011.QC.jpg
4d9d6b3b00e827df7840603fbc3d1285
ec53a1c97ecba7fc670970b90eda7409c47898bc
1860 F20101130_AABXRM fickett_f_Page_132.txt
d6717f1a37f1a05e27da373999061165
3f3f697828d11e92380aa56377c74f1e7df66dd3
1740 F20101130_AABXQY fickett_f_Page_096.txt
56e845372feb9c1de13701c355419c32
ead0127315afc5be63b3f576cbfb9fc29ab2e899
1996 F20101130_AABWNW fickett_f_Page_036.txt
8ab6b4841f955b5c0ac42b81a5f0a086
695d012209d8957a6a7f009fb03394b91eadad46
1842 F20101130_AABWOK fickett_f_Page_145thm.jpg
a6481794f9bdebad843f8921e336c62c
fbd074e26d94597086ed173fdb176d87400d4b4c
3630 F20101130_AABXSB fickett_f_Page_011thm.jpg
d9fd00c2321493335b1ad61918bcd591
6f1ebe7ac38130cb64b129172aa66c53c44c895f
2593 F20101130_AABXRN fickett_f_Page_137.txt
3c781a9d74c73984f3fbeaac49e58277
a6388e2798a30ab51b06e6cc9f83f1b67e52796b
1814 F20101130_AABXQZ fickett_f_Page_098.txt
fea9f5e6e93b944019f811608a466c45
3bd4dd810c0fbcf37484e35811c331d1374b4d25
F20101130_AABWNX fickett_f_Page_026.tif
eb98462b3840380de04bcd2e44fe9338
3cbe4afd6bc6b763a4a77443ce9dd67c247d1e84
64777 F20101130_AABWOL fickett_f_Page_054.jp2
577ae44c0c29713033ddede4985fa775
6feb298aea20ec4d0671e522df40cc7020e6c29c
5268 F20101130_AABXSC fickett_f_Page_012thm.jpg
33a9628c96b5849883dd83838c35786a
a840aec880b6a3f96ff17c9dacba7cb35e80f19f
2799 F20101130_AABXRO fickett_f_Page_140.txt
336f93712606bdbe5339f739cde8de20
49307f662549c479e3b0da23234e601584fc767f
59917 F20101130_AABWNY fickett_f_Page_096.jp2
cc6483a10b40b95bd3584ce76e72811d
d40db4dc63e208b14167eab1ea1e6dabdfac403d
1968 F20101130_AABWPA fickett_f_Page_075.txt
7ae006ee7aa0cb9c6cdbc1ea23a2129d
7b1b8c67492c615fdf40c222e18a684fadd5c40d
F20101130_AABWOM fickett_f_Page_064.tif
1b55a45e5ff967d4e4501203744f5d41
b296074d9af40672b7b0b420367e2160b0bb72ea
23940 F20101130_AABXSD fickett_f_Page_013.QC.jpg
0887c4204181a7f23b6d8280cb34802e
275bacbb6bc7eb2d4948a2da2e43102205dffc7c
2664 F20101130_AABXRP fickett_f_Page_142.txt
11014fd06a5fc700c6a73467073700f7
488a73e991f58c35bdfd2849c8928dc6317789d7
14317 F20101130_AABWPB fickett_f_Page_095.QC.jpg
d7c20c950677d1f3ce8eec804012208a
6af5c191f5ea25a8b80c5b08a36febcfc71f3adc
6690 F20101130_AABWON fickett_f_Page_084thm.jpg
2df50dd05499846bfbb128a7bb7f01ee
b258164258ccfba933d9d3ae0ceaeb302615338b
21061 F20101130_AABXSE fickett_f_Page_014.QC.jpg
750aae9f415fa813f725224062fd65a6
f12557a9ac6235db3aac534ea48821a365295484
2693 F20101130_AABXRQ fickett_f_Page_144.txt
d41c393fa25d27e504603e67137aa301
1eb8daa1945ddb04556c108ee508ca8c9cabc7f4
62307 F20101130_AABWNZ fickett_f_Page_009.pro
6c9e55ef0b0b492a3926f1317d80c763
5bbefc4b485f98709ecd8fbf77fa99232aafb095
27732 F20101130_AABWPC fickett_f_Page_096.jpg
cb9c590eb570607d6ff1d8e948b956b9
e532207d306f576f13be0bee3b30ad1c853c678a
48224 F20101130_AABWOO fickett_f_Page_068.pro
56d9f22b07ef0b0f529a9c661d935ba8
90caf31a7cfbe2edb6441a6d57594524669a141d
6041 F20101130_AABXSF fickett_f_Page_014thm.jpg
cf51bd3be2ac54edff1369167eff810c
1ba427de9c4ded028202088c9e5eaf4a111b7bd8
8098 F20101130_AABXRR fickett_f_Page_001.QC.jpg
a0160d53c5b5f5144bfd84ed0e47c15e
749ae85b69011b2ed7c99fd0ea47a39127e18b4b
741 F20101130_AABWPD fickett_f_Page_107.txt
2385d4aa6c662dfa8f24193efcde265c
1146a7b28e463d0f12e97445aa40d6c632a86e31
4913 F20101130_AABWOP fickett_f_Page_112thm.jpg
3af05557ca711914ab2785306e06aa65
8913e8a77fddfd32b506a27dee3375053fb3aedf
6415 F20101130_AABXSG fickett_f_Page_015thm.jpg
edc81ea81f88886bc4a821e72aca2ec7
6bf9e70bf4855d2130613e57e66f51553cc6ca6a
2595 F20101130_AABXRS fickett_f_Page_001thm.jpg
29faf383ee130830fc2f22669f8bb484
7a0fa12fe1662e181ac16ce1553cdcd01681de2c
3130 F20101130_AABWPE fickett_f_Page_122thm.jpg
8af3e8c433fe3d3261219fe1dc2c8fc8
f28084c0c75481ff1de71edf1625cad06465e557
73511 F20101130_AABWOQ fickett_f_Page_023.jpg
37eae391a4718bad393751301673b899
f86f1a3aa5902888f6c84a58afea9ec5adb74be4
F20101130_AABXSH fickett_f_Page_016.QC.jpg
c3c6ee079b65609e01d7ee8803ec7f03
269a5ecc077b2384d0a20c6e13a9e840847073c9
3307 F20101130_AABXRT fickett_f_Page_002.QC.jpg
7338b37e7311eef84c7e375a853f2493
6443ecd76903c0a878aa2687a085b792eeb8e929
93683 F20101130_AABWPF fickett_f_Page_058.jp2
f7f899bb3a292626b6078d4dc0f263ec
9e1beb9ccc215d40d1368bd3b3f0c7ff721a85fd
2975 F20101130_AABWOR fickett_f_Page_098thm.jpg
16eb4471271b75c1858a752d93d31cc6
a4268a84101d6fdf360a55d2fb9e23fb841ecf02
5266 F20101130_AABXSI fickett_f_Page_016thm.jpg
1d6336e5806086a9fb8bb26b4a4b9677
4aff6de55749bae30687822c240d6555d1f0a982
1379 F20101130_AABXRU fickett_f_Page_002thm.jpg
64d677f3c39b36b6a6390217b955f9d8
d054fe0fcf4e08333108be79b840efc663de16d0
90027 F20101130_AABWPG fickett_f_Page_142.jpg
08262e4efb0394eaf4176952363fb63f
ae9a90ea7d502d7f15731ae9944a9da71775d324
F20101130_AABWOS fickett_f_Page_099.tif
608d96d455692d2a86d7a0f5853f2910
67cd8977c57a2aa0e7c8ce5b999e8fdf5c3aeedf
20138 F20101130_AABXSJ fickett_f_Page_017.QC.jpg
b5aa740d8d99b3aca71c5a46134989ab
764fce7acb19b0662ec97d20c68b5f89f50a3148
6171 F20101130_AABXRV fickett_f_Page_004thm.jpg
7588876c691972d0622afc7b25ca98c1
7144a9bca05d249796cc6336ff61f09cf32d3c7e
23712 F20101130_AABWPH fickett_f_Page_035.QC.jpg
32a6a0047d42f9948967c46a20007cbc
28bc1f0a8b9e82e1884930301d31886e78973c47
2523 F20101130_AABWOT fickett_f_Page_133thm.jpg
8b1dde6418603fd80ab44d6dc395d61b
e3c85deb3b588e2e23643f85209ab181f391c08f



PAGE 1

EFFECTS OF THE TIMING OF INITIA TION OF FAT SUPPLEMENTATION ON PRODUCTIVE AND REPRODUCTIVE RESPONSES OF PERIPARTURIENT DAIRY COWS DURING SUMMER By FAITH MARIE CULLENS A THESIS PRESENTED TO THE GRADUATE SCHOOL OF THE UNIVERSITY OF FLOR IDA IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE UNIVERSITY OF FLORIDA 2005

PAGE 2

Copyright 2005 by Faith M. Cullens

PAGE 3

This thesis is dedicated to my wonderful and patient husband, Gus, and to my mom who has always encouraged me to pursue my dreams.

PAGE 4

iv ACKNOWLEDGMENTS I would like to thank my committee, Dr. Charles Staples, Dr. Lokenga Badinga, and Dr. William Thatcher for their time and dedication to my education. I would like to thank the Davis family fo r reoccurring scholarship monies, as well as the deans office for matching my assist antship. In addition, the IFAS, Graduate Student Association, and The Animal Scien ces Graduate Student Association travel grants were a great financial assistance in traveling to the American Dairy Science Association Annual meeting to present my research. I would also like to thank Catarina Silveria and Myriam Lopez for all of their help with animal work and with data entry. I w ould like to thank Todd Bilby, Flavio Silvestre, Julian Bartolome, Alessandro Sozzi for their help with reproductive management of the cows. Joyce Hayen played an essential role in pre-trial advising and gave extremely valuable assistance with lab work. Trem endous help with sample collection and technical assistance was given by my fr iends and co-workers including Lucia Holtshausen, Bruno Amaral, Carlos Alosilla, Marcio Liboni, Tomas Belloso, Sergei Sennikov, Wimberley Krueger and Elizabeth Johnson. I also would like to thank Dr. Raymon Littel for his valuable statistical support and Dr. Charles Wilcox for dedicating many lunch hours to assisting me with statis tics. I also would like to thank Dr. John Arthington for his help analyzing acute pha se proteins and Dr. Duane Keisler for analyzing plasma leptin concentr ations. Last but not least, I must thank Mary Russell and the DRU staff who helped with animal work.

PAGE 5

v TABLE OF CONTENTS ACKNOWLEDGMENTS.................................................................................................iv LIST OF TABLES............................................................................................................vii LIST OF FIGURES...........................................................................................................ix ABBREVIATIONS...........................................................................................................xi ABSTRACT......................................................................................................................x ii CHAPTER 1 INTRODUCTION........................................................................................................1 2 LITERATURE REVIEW.............................................................................................4 Fatty Acids Defined......................................................................................................4 Effects of Supplemental Fat on Feed Intake and Production.......................................5 Timing of Initiation of Fat Feeding............................................................................10 Effects of Supplemental Fat on Adipose Tissue Lypolysis........................................12 Effects of Supplemental Fat on Immune Function.....................................................15 Effects of Supplemental Fat on Reproduction............................................................17 A Historical Review............................................................................................17 The Modern Dairy Cow.......................................................................................18 Effects of Supplemental Fat on Follicle Development...............................................24 3 EFFECTS OF THE TIMING OF IN ITIATION OF FEEDING CALCIUM SOAPS OF LONG CHAIN FATTY ACIDS ON PERIPARTURIENT HOLSTEIN COWS DURING SUMMER.................................................................45 Materials and Methods...............................................................................................45 Cows and Diets....................................................................................................45 Sample Collection...............................................................................................46 Reproductive Management..................................................................................47 Animal Health.....................................................................................................49 Sample Analysis..................................................................................................49 Statistical Analysis..............................................................................................52

PAGE 6

vi Results and Discussion...............................................................................................54 Dry Matter Intake and Diets................................................................................54 Production and Body Weight..............................................................................55 Plasma Metabolites..............................................................................................57 Plasma Hormones................................................................................................60 Reproductive Measurements...............................................................................66 Hepatic Measurements........................................................................................70 Measurements of Immune Status........................................................................72 Parity Effects.......................................................................................................76 Conclusion...........................................................................................................77 APPENDIX TESTS OF HETER OGENITY OF REGRESSION..................................116 LIST OF REFERENCES.................................................................................................121 BIOGRAPHICAL SKETCH...........................................................................................133

PAGE 7

vii LIST OF TABLES Table page 2.1 Effects of feeding ruminally inert fat (C a salts of fatty acids (CSFA) or prilled fat (PF)) to the same cows throughout the study on DMI, milk production, milk composition, and body weight (BW) change...........................................................27 2.2 Effects of feeding oilseeds alone or in combination with other fat sources to the same cows throughout the experiment on DMI, milk production, milk composition, and body weight (BW) change...........................................................33 2.3 Effects of feeding rendered fats to the same cows throughout the study on DMI, milk production, milk composition and body weight (BW) change........................37 2.4 Effects of feeding ruminally inert fat (C a salts of fatty acids (CSFA) or prilled fat (PF)) to the same cows throughout the study on concentration of plasma hormones and metabolites........................................................................................40 2.5 Effects of feeding oilseeds alone or in combination with other fat sources to the same cows throughout the study on concentration of plasma hormones and metabolites...............................................................................................................43 2.6 Effects of feeding rendered fats to the same cows throughout the study on concentration of plasma hormones and metabolites................................................44 3.1 Ingredient and chemical composition of diets fed to nonlactating cows.................78 3.2 Ingredient and chemical composition of diets fed to lactating cows.......................79 3.3 Milk yield, milk composition, post partum body weight, and postpartum body condition score.........................................................................................................80 3.4 Concentration of plasma hormones and metabolites and day of first ovulation......81 3.5 Concentration of plasma PGF2 metabolite (PGFM) the first 14 DIM and the size and characteristics of the uterus and cervix......................................................82 3.6 Vaginal observations at 21 and 28 DIM...................................................................83 3.7 The number and size of ovarian structures at 21 and 28 DIM.................................84 3.8 Ovarian structures present on the ovaries................................................................85

PAGE 8

viii 3.9 Concentrations of hepatic triacylglycer ol (TAG), and hepatic IGF-I, IGF-II, and IGF binding protein (BP) -2 mRNA levels..............................................................86 3.10 Concentration of plasma acute ph ase proteins and liver enzymes...........................87 A-1 Tests of homogeneity of regression and orthogonal contrasts for treatment effects on plasma Insulin, IGF-I and accu mulated progesterone concentrations...116 A-2 Tests of homogeneity of regression and orthogonal contrasts for treatment effects on plasma PGF2 metabolite (PGFM), haptoglobin, and ceruloplasmin concentrations.........................................................................................................117 A-3 Tests of homogeneity of regression and orthogonal contrasts fo r parity effects on plasma insulin, IGF-1, accumulated progesterone, PGF2 metabolite (PGFM), haptoglobin, and ceruloplasmin concentrations.....................................................118 A-4 Tests of homogeneity of regression a nd orthogonal contrasts for treatment by parity interaction effects on plasma insulin, IGF-1 and accumulated progesterone concentrations...................................................................................119 A-5 Tests of homogeneity of regression a nd orthogonal contrasts for treatment by parity interaction effects on plasma PGF2 metabolite (PGFM), haptoglobin, and ceruloplasmin concentrations..........................................................................120

PAGE 9

ix LIST OF FIGURES Figure page 3.1 Least squares means for milk production.................................................................88 3.2 Least squares means for body weight......................................................................89 3.3 Least squares means for body condition score.........................................................90 3.4 Least squares means for plasma BHBA concentration............................................91 3.5 Least squares means for pl asma NEFA concentration.............................................92 3.6 Least squares means for plas ma glucose concentration...........................................93 3.7 Least squares means for blood urea nitrogen (BUN) concentration........................94 3.8 Least squares means for plasma leptin concentration..............................................95 3.9 Regression plot of plasma IGF-1 c oncentration of primiparous (A) and multiparous (B) cows .............................................................................................96 3.10 Regression plot of plasma IGF-1 c oncentration of primiparous (A) and multiparous (B) cows...............................................................................................97 3.11 Regression plot of plasma insulin concentration......................................................98 3.12 Regression plot of accumulated plasma progesterone concentration from d 1 to 77 postpartum...........................................................................................................99 3.13 Regression plot of accumulated plasma progesterone concentration from d 1 to 46 postpartum.........................................................................................................100 3.14 Regression plot of conc entration of plasma PGF2 metabolite..............................101 3.15 Least squares means for hepatic triacylglycerol concentration..............................102 3.16 Ten micrograms of total cellular RNA is olated from livers of cows fed diets without CSLCFA (lanes 1 to 6), or diet s with CSLCFA starting prepartum (lanes 7 to 12) or at 1 DIM (lanes 13 to 18) were subjected to Northern blot analysis. Representitive Northern blots for IGF-I (A), IGF-II (B), or IGFBP-2 (C) mRNA expression are shown.............................................................................................103

PAGE 10

x 3.17 Least squares means for he patic IGF-I mRNA abundance....................................104 3.18 Least squares means for he patic IGF-II mRNA abundance...................................105 3.19 Least squares means for hepa tic IGFBP-2 mRNA abundance..............................106 3.20 Regression plot of plasma concentration of haptoglobin.......................................107 3.21 Regression plot of plasma c oncentration of ceruloplasmin...................................108 3.22 Least squares means for plasma c oncentration of total biliruben..........................109 3.23 Least squares means for plasma concen tration of alanine aminotransferase.........110 3.24 Least squares means for plasma alkaline phosphatase...........................................111 3.25 Least squares means for plasma concen tration of aspartat e aminotransferase......112 3.26 Least squares means for plasma concentr ation of gamma glutamyl transferase....113 3.27 Least squares means for concen tration of plasma albumin....................................114

PAGE 11

xi ABBREVIATIONS AI artificial insemination ALK alkaline phosphatase ALT alanine aminotransferase AST aspartate aminotransferase BW body weight BCS body condition score BHBA -hydroxy butyric acid BUN blood urea nitrogen CSFA calcium salts of fatty acids CSLCFA calcium salts of long chain fatty acids CL corpus leutem CLA conjugated linoleic acid CV coefficient of variation DM dry matter DIM days in milk DMI dry matter intake EE ether extract FA fatty acids FCM fat corrected milk FSH follicle stimulating hormone GGT gamma glutamyl transferase GnRH gonadatropin releasing hormone LH leutinizing hormone LA linoleic acid LNA linolenic acid NEFA nonesterified fatty acids NFC non fiber carbohydrates PGF2 prostaglandin F2 PGFM 13, 14-dihydro-15-keto-PGF2 metabolite PHT partially hydrogenated tallow PUFA polyunsaturated fatty acids RIA radioimmuno assay RFM retained fetal membranes SCC somatic cell count TAG triacylglycerol TMR total mixed ration WCS whole cottonseed

PAGE 12

xii Abstract of Thesis Presen ted to the Graduate School of the University of Florida in Partial Fulfillment of the Requirements for the Degree of Master of Science EFFECTS OF THE TIMING OF INITIA TION OF FAT SUPPLEMENTATION ON PRODUCTIVE AND REPRODUCTIVE RESPONSES OF PERIPARTURIENT DAIRY COWS DURING SUMMER By Faith M. Cullens May 2005 Chair: Charles R. Staples Major Department: Animal Sciences Primiparous (n = 22) and multiparous (n = 25) Holstein cows were utilized in a completely randomized block design to determin e the effects of timing of the initiation of feeding Ca salts of long chain fatty acids ( CSLCFA ) on cow performance the first 100 days in milk ( DIM ). Four treatments were as follows: control (no CSLCFA) and CSLCFA supplementation (2% of dietary DM ) beginning at 28 d prior to expected calving date (CSLCFA prepartum), at 1 DI M, or at 28 DIM. Cows fed CSLCFA beginning in the prepartum pe riod tended (P > 0.05 but 0.10) to produce more milk (42.2 vs. 37.1 kg/d) and had (P 0.05) fewer small (2 to 5 mm) ovarian follicles during the first estrous synchronization using injec tion of gonadotropin-releasing hormone (44 3 DIM) than cows fed CSLCFA initiated after calving. This milk increase was accompanied by a lower incidence of disease (mastitis, metritis, or retained fetal membranes) in the first 10 DIM (8 vs. 43%), el evated concentrations of triacylglycerol in

PAGE 13

xiii the liver dry matter ( DM ) at 14 2 DIM (23.0 vs. 10.4%), lower expression of hepatic IGF binding protein-2 mR NA (0.08 vs. 0.12 arbitrary un its/18S), and elevated concentrations of plasma b ilirubin (0.36 vs. 0.23 mg/dl) compared to cows not fed CSLCFA prepartum. Expression of hepa tic IGF-I and IGF-II mRNA did not differ among treatment groups. Multiparous cows appeared to benefit more from supplementation with CSLCFA than primiparous cows in that multiparous cows not fed CSLCFA at any time during the study had or tend ed to have lower concentrations of milk protein, a longer period and greater lo ss of body weight postpartum, greater concentrations of plasma beta-hydroxy butyr ic acid, and lower concentrations of plasma glucose, leptin, and IGF-I. In addition, multiparous cows fed fat prepartum tended to have fewer small (2 to 5 mm) and medium (6 to 9 mm) size but more larger size ovarian follicles ( 10 mm in size) at 21 DIM, more uterin e tone at 21 DIM, a smaller cervical os at 28 DIM, and a slower decrease in plas ma concentrations of 13, 14 dihydro-15 keto prostaglandin F-2 metabolite the first 10 d postpartum than multiparous cows not fed fat prepartum. Feeding CSLCFA in the prepar tum period also benefited primiparous cows in that they tended to have lower concentra tions of plasma fibrinogen. Initiation of fatfeeding at calving tended to result in fe wer and smaller corpus luteum at 21 DIM compared to those fed CSLCFA prepartum. Estimated day of first ovulation postpartum based upon two consecutive days of proge sterone exceeding 1.0 ng/ml did not differ among treatment groups (mean of 27 DIM). Multiparous cows fed CSLCFA had greater concentrations of plasma IGF-I at the time of artificial insemination and conception rate at first insemination tended to be bette r for all animals fed CSLCFA (58 vs. 27%) regardless of day of initiati on of CSLCFA supplementation.

PAGE 14

1 CHAPTER 1 INTRODUCTION The transition from pregnancy to lactation in the dairy cow is a cr itical phase of the lactation cycle. The transition period, typically considered the time three weeks prepartum until three weeks postpartum, is marked by declining dry matter intake ( DMI ) as the cow approaches parturition and negativ e energy status in early lactation. Total energy intake in early lactation is usually le ss than what is required for maintenance and milk production (Staples et al., 1990); therefor e the cow must mobilize adipose stores in the form of nonester ified fatty acids ( NEFA ) to help support lactation. Nonesterified fatty acids are removed from circulation by th e liver. Once in the liver, NEFA are utilized for energy; however ruminants have reduced ability to export triglycerides from the liver and lipids can accumulate causing a metabolic disorder known as fatty liver (Drackley, 1999). Supplemental fats can increase the energy de nsity of the diet and reduce an energy deficit in early lactation. However milk pr oduction often increases when fats are added to the diet, resulting in no improvement of energy status. In a ddition, fat feeding can often result in a depression of DMI (Allen, 2000). The mechanisms by which supplemental fat depresses DMI are not appare nt but could involve negative effects on ruminal fermentation and gut motility, release of gut hormones, oxidation of fat in the liver, and palatability of di ets containing added fat (Alle n, 2000). Feeding supplemental fat in the form of Ca salts of fatty acids ( CSFA ) makes the fat partially inert in the rumen and sometimes can prevent a depression in DM I. Feeding CSFA can allow specific fatty

PAGE 15

2 acids ( FA ) to escape ruminal biohydrogenation whic h are then available in the small intestine for absorption and utilization. The modern dairy cow may be deficient in the essential FA, linoleic acid ( LA ) (Sanchez and Block, 2002). Providing LA in the form of CSFA may reduce a deficiency and act on targ et tissues in addition to increasing the energy density of the diet. Supplemental fats have proven beneficial to reproductive efficiency of lactating dairy cows as well (Staples et al., 1998). Although reproductive perf ormance is strongly associated with energy status (Staples et al ., 1990), dietary fats can provide FA precursors for cholesterol and prostaglandin production, which have an affect on ovarian function, uterine function, and conception rates. Immune reactions have also been shown to be modulated by the diet including the FA composition by influencing cellular communication and activation th rough the synthesis of pros taglandins (Calder et al., 2002). Decreased incidence of disease in ea rly lactation can resu lt in increased milk production throughout the lactation and an incr ease in reproductive efficiency. The amount of a particular FA (e.g., LA) stored in the target tissue may control prostaglandin synthesis. The modern dairy cow in the United St ates is most pr oductive between the temperatures of 5 and 15C and will begin to experience slight lo sses in milk production between 15 and 25C (Hahn, 1985). Above 26 C, dramatic losses in production can occur although the humidity index will alter th is upper critical temperature (Berman et al., 1985). In an attempt to dissipate additional body heat, the cow will increase her respiration rate, increase sweating rate, incr ease blood flow to skin, and decrease energy intake (Blackshaw and Blackshaw, 1994). Because fat has a low heat increment

PAGE 16

3 associated with feeding, has a high energy dens ity, and is utilized with high efficiency, it is an ideal feed additive duri ng periods of heat stress. Re ducing heat stre ss in lactating dairy cattle can increase milk production a nd reproductive efficienc y. Heat stress can alter hormonal profiles and re duce the duration and intensity of estrus (Wolfenson et al., 1988). Using a large group of Isreali cows, Zeron a nd coworkers (2001) reported lowered conception rates in summer versus wi nter months which may have been due to differences in follicular dynamics and changes in the biological membranes. In the winter, more follicles per ovary and embryo development to the blastocyst stage, the twoto four-cell stage, and the morula stage we re improved compared to summer months. In addition, the proportion of FA in the phospho lipids of oocytes, granulosa cells, and follicular fluid was increased in the summer whereas the proportion of polyunsaturated FA in the phospholipids of oocytes and granul osa cells was increased in the winter. A study examining pregnancy rates in Florid a and Georgia herds using DHIA records reported that high milk production exacerbated th e already great drop in non-return rates during the summer months (A l-Katanani et al., 1999). The objective of this thesis was to evaluate the timing of initia tion of Ca salts of long chain FA on the production, plasma hor mones and metabolites, and reproduction of periparturient Holstein cows in summer.

PAGE 17

4 CHAPTER 2 LITERATURE REVIEW Fatty Acids Defined Linoleic acid ( LA ), the major fatty acid ( FA ) in most oilseeds, and linolenic acid ( LNA ), the major FA in fresh forages, are considered essential FA because they cannot be synthesized by mammals or ruminal microorganisms. They lack the 12 and 15 desaturase enzymes to convert oleic acid to LA and LNA. Both LA and LNA are polyunsaturated FA ( PUFA ) in that they have more than one point of unsaturation, or double bond. The shorthand notation system fo r identifying FA includes a description of the number of carbons present within the chain, the number of double bonds, and the location and configuration of the double bonds. In addition, the -numbering system begins numbering carbons starting at the met hyl end of the FA. For example, LA is notated as C18:2, -6 because it has 18 carbons, tw o double bonds, and the first double bond is at the sixth carbon from the met hyl end. The LNA is designated C18:3, -3. Lastly, FA can be classified in the -9 category such as oleic acid, C18:1. Omega families cannot be interconverted. Fat supplementation is commonly used to incr ease the energy density of the diet of lactating dairy cows. During the first few w eeks of lactation, dairy cows are limited by nutrient intake by which to meet the demands of lactation. Earl y postpartum dry matter intake ( DMI ) is limited by ruminal fill and does not peak until 10 to 14 wk postpartum, while milk production usually peaks at 4 to 8 wk postpartum (Nati onal Research Council [NRC], 2001).

PAGE 18

5 Feeding diets with 3 to 5% added fat can increase energy intake without having a major negative effect on fiber digestion or milk fat (Palmquist and Jenkins, 1980). Supplying energy in the form of fat in place of carbohydrate can reduce microbial protein production since carbohydrates ar e the primary energy source for ruminal microbes. On the other hand, increasing the energy concentra tion of the diet by increasing the starch to an excessive concentration can have detrimen tal effects on digestion and animal health. Feeding excessive unsaturated FA, which ha ve a toxic effect on ruminal microbes, may lead to depressed fiber digestion a nd milk fat production (Brooks et al., 1954; Jenkins and Palmquist, 1984). Manufact uring Ca salts of fatty acids ( CSFA ) can make them partially inert in the rumen and can increase the energy density of the diet without hindering forage digestion (Je nkins and Palmquist, 1984). Pr eformed soaps of FA should not dissociate in the rumen, but dissociate in th e abomasum at a low pH. The FA are then available in the small intes tine for absorption and utiliza tion by the body (Jenkins and Palmquist, 1984). The studies reviewed in this thesis have been limited to those in which cows were fed the same experimental diets for the durati on of the study. Importa nt aspects of each study are summarized in Table 2.1 (ruminally in ert fats), Table 2.2 (oilseeds) and Table 2.3 (rendered fats). Effects of Supplemental Fat on Feed Intake and Production Dry matter intake is often affected by the addition of fat to the diet (Allen, 2000; Jerred et al., 1990) The type of fat fed as we ll as the type and amount of forage will have an effect on the extent to which DMI is affected (Allen, 2000). Although the mechanisms are unclear, intake may be depressed when supplemental fats are fed due to decreased palatability, ruminal fill due to inhibition of fiber digestion, the metabolic

PAGE 19

6 regulation of the gut hormone cholecystoki nin on the brain satiety centers, and an increased rate of FA oxidation in the liver that can alter signals generated by hepatic vagal afferent nerves to br ain centers signaling satiety (Allen, 2000). In addition, PUFA that reach the small intestine may decreas e gut motility which could decrease DMI (Drackley et al., 1992). In a review of seve ral studies, Allen (2000) reported that there was a linear decline in DMI such that for ev ery 1% inclusion of tallow or CSFA in the diet there was a reduction of DMI of 1.2 and 2.5% respectively. Allen also reported that oilseeds negatively affected intake, although in a quadratic fashion with 2% of dietary FA coming from oilseeds resulting in maximum intake suppression. In this review of studies where CSFA we re fed to the same animals throughout the study (Table 2.1), the majority of experiment s report no effect on DMI when CSFA were fed at 1.8 to 5% of dietary dry matter ( DM) (Atwal et al., 1990; Chouinard et al., 1997; Erickson et al., 1992; Firkins and Eastridge, 1992; Holter et al., 1992; Moallem et al., 2000; Palmquist and Weiss, 1994; Schroeder et al., 2003; Skaar et al., 1989; Spicer et al., 1993). However, several studies reported a depression in DMI when cows were fed CSFA in diets in which alfalfa (silage or ha y) was the sole forage source (Chouinard et al., 1997; Harrison et al., 1995; Jerred et al., 1990; Simas et al., 1995). Interestingly, a few studies reported DMI was depressed early in the experiment; however the effect disappeared after the cows consumed the diets for a longer period of time (Beam and Butler, 1998; Chouinard et al., 1997; Garcia-B ojalil et al., 1998), suggesting that there may be an adaptation period. Depression of DMI was less commonly repor ted when cows were fed oilseeds as a fat source throughout the durat ion of the study. In two studies by Harrison and

PAGE 20

7 coworkers (1995), intake was depressed when early lactation cows consumed diets of 12% whole cottonseed ( WCS ) and 2.7 or 5% of DM as CSFA (Table 2.2). In one of those studies, DMI was depressed when a diet of 12% WCS without CSFA was fed. When high amounts of whole soybeans (18% of dietary DM and 6.2% dietary ether extract ( EE )) were fed in a diet c ontaining 30% corn silage an d 20% alfalfa silage, DMI was depressed 2.3 kg/d (9.7% of DMI) in comparison to co ntrols (3.2% dietary EE). However the feeding of soybeans in the gr ound roasted form (18% of dietary DM) did not affect DMI (Pires et al., 1996). The roasting process may reduce exposure of the oil to the rumen microbes and th erefore reduce the negative in fluence on rumen function. Escape of LA to the small intestine was increased in cows fed roasted versus raw soybeans (Dhiman et al., 1995; Tice et al., 1994). Similarly, the addition of rolled safflower seeds at 10% of dietary DM in co mbination with bST injections (5.5% dietary EE) depressed DMI by 4.6 kg/d (19.2% of DMI) in comparison to control cows receiving bST injections (2.3% dietary EE) in a diet of 25% corn si lage and 25% alfalfa hay (DM basis). However, the replacement of safflower seeds with rolled sunflower seeds (6.2% dietary EE) in combination with bST injec tions did not affect DMI (Stegeman et al., 1992). In this review of studies where rendere d fats were fed to the same animals throughout the experiment, only two studies reported a suppression in DMI in comparison to the control or similar treatme nt without fat (Table 2.3). Bateman and coworkers (1996) reported a depression of 2.6 kg/d (10% of DMI) when tallow (2% of dietary DM) was fed in a 33% NDF diet in the winter (3.5 and 5.4% dietary EE for fatsupplemented and control diets, respectively) However, tallow feeding had no effect on

PAGE 21

8 DMI when included in the summer or in a di et with 40% NDF. Lastly, when lactating cows were fed diets of 0 or 3% tallow (DM ba sis) in a ration comprised of 33% alfalfa haylage and 17% corn silage, DMI was de pressed 0.9 kg/d (3.6% of DMI) and 1.3 kg/d (5.4% of DMI) with the addition of 0 or 5% escape protein supplement, respectively (Son et al., 1996). Changes in milk production can accompany changes in DMI. Often, milk production is increased due to increased energy intake when supplemental fats are fed, particularly in early lactation when the da iry cow must depend on her body reserves to help satisfy energy requirements for maintena nce and lactation. Of 28 studies in which ruminal inert fat was fed to cows in conti nuously applied treatments eleven studies did not report an increase in production of milk or fat-corrected milk ( FCM ) (Table 2.1). Reasons for the discrepancies between studies may be due in part to the day postpartum that fat feeding was initiate d, the duration of the study, f eed quality, amount of milk production, and combinations of these factor s. In one study that did not report an increase in milk production, cows were suppl emented CSFA while on pasture (Schroeder et al., 2003). In another st udy that did not report an incr ease in milk production, a 0.45 kg drench of CSFA was administered for only 4 d postpartum (Pickett et al., 2003). Cervantes and coworkers (1996) did not repor t an increase in milk production when feeding 0.4 kg/d of CSFA but the cows were in midlactation (average of 112 DIM) and were only fed diets for 38 d. Additionally, two studies that did not report an increase in milk production were already feeding high conc entrate diets (> 61% of dietary DM) that included > 8.5% WCS and the additional en ergy provided by dietar y fat may not have been beneficial (Simas et al., 1995; Spicer et al., 1993).

PAGE 22

9 Similarly, in 11 of 17 studies in which oilseeds were fed to the same animals throughout the experiment, milk production or FCM increased (Table 2.2). Two studies that did not report an increase in milk produc tion were already f eeding high concentrate diets (> 60% of dietary DM) (Khorasani et al., 1991; Markus et al ., 1996). Another study reported DMI depression of 9.7% when w hole soybeans were fed, which may have contributed to the unaffected milk production (Pires et al., 1996). When rendered fats were fed in continuous studies, six of 13 studies reported an increase in milk production or FCM yield (Table 2.3), and intere stingly there was no suppression in DMI. Reasons for this vari ation in response across studies were not apparent. Milk protein concentration was commonly re ported to decline when ruminally inert fats or oilseed were fed (Tables 2.1 and 2.2) but less often when rendered fats were fed (Table 2.3). Although the mechanism is unc lear, it could be due, in part, to less glucogenic precursors being consumed as starch is replaced with lipid. Less glucogenic precursors present in the diet is associated with decreased milk protein concentration (Rigout et al., 2003). The response of milk fat concentration to supplemental fat seems to be dependent upon many factors. About 50% of the fat found in milk is synthesized in the mammary gland from acetate and butyrate, while the ot her 50% comes directly from fat absorbed from the blood (Ackers, 2002). Palmquist and coworkers (1993) published an equation predicting an increase in milk fat concentration of 0.18% fr om feeding an additional 0.5 kg of fat daily. Conversely, milk fat depression is often seen when fat is fed in diets in which corn silage was the sole forage s ource, resulting in a mo re acidic ruminal

PAGE 23

10 environment than when fed in combination with alfalfa hay or haylage which has a greater buffering capacity (Onetti et al., 2004; Ruppert et al., 2003; Smith et al., 1993). This is thought to occur when PUFA are biohydrogenate d in the rumen under acidic conditions to trans C18:1 FA rather than to C18:0. The enzymes responsible in the mammary gland to synthesize the short and me dium chain fatty acids found in milk may be inhibited by these trans C18:1 FA, especially trans -10 C18:1, resulti ng in decreased milk fat concentration (Bauman and Griinari, 20 03). In this review of studies in which supplemental fat was fed to the same cows throughout the experiment, an increase in milk fat concentration was more consistent when cows were fed CSFA in comparison to oilseeds or rendered fats. Accompanying an increase in milk produc tion due to fat supplementation may be an increase in 4% FCM yield (Erickson et al ., 1992; Moallem et al ., 2000; Salado et al., 2004). However, if milk fat concentra tion is unchanged or is depressed by fat supplementation, yield of 4% FCM may not signi ficantly increase despite an increase in milk production (AbuGhazaleh et al., 2004; Pa ntoja et al., 1996; Selberg et al., 2004). Timing of Initiation of Fat Feeding Few studies in the literature examine when to start feeding fat during the periparturient period. If fat supplementation is begun in the dry period, benefits can be expected due to the adjustment of the animal s palate and ruminal microflora before the onset of the lactation. Skaar and workers (1989) fed 40 multiparous cows diets of 0 or 5% prilled fat containing 50% forage (equal amounts of corn silage and alfalfa silage, DM basis) beginning at 17 d be fore expected calving date and continued through 105 d in milk ( DIM ). Intake did not differ between fatfed and control cows yet overall milk production tended to increase for cows fed fa t. During the cool season, milk production

PAGE 24

11 did not differ between treatments, however, dur ing the warm season milk yield increased from 34.5 to 44.3 kg/d. Although cow performa nce was not negatively affected, liver biopsies revealed that cows fed fat beginni ng prepartum tended to have increased total hepatic lipid concentrations in comparison to controls (27.5 vs. 26.1%, DM basis at 1 DIM and 29 vs. 24%, DM basis at 5 weeks postp artum). Plasma nonesterified fatty acid ( NEFA ) concentrations did not differ between treatments indicating that the additional lipid in the liver may have been from dietary origin. Waiting until calving to start feeding fat is another approach often tried because of the extra costs associated with fat supplementation. University of Florida workers (Garcia-Bojalil, et al., 1998) fed multiparous cows corn silage/alfalfa hay-based diets containing 0 or 2.2% calcium salts of palm oil beginning at calving and continuing through 120 DIM. Fat-fed cows began to produc e more milk (~2 kg/d) after 3 weeks of supplementation and continued to produ ce more milk throughout the experiment. Allowing cows to get through the period of negative energy balance before adding fat to the diet is one more strategy employed. Schingoethe and Casper (1991) summarized five studies in which whole sunflower or extruded soybeans were fed beginning at 4 weeks and continued through 16 weeks of lactation. Cows produced an average of 0.9 kg/d more milk while consuming the seeds, however, there was still a 3 to 4 week delay before a m ilk response was evident. Salfer and coworkers in Minnesota (1995) ev aluated when to initiate the feeding of partially hydrogenated tallow ( PHT ). Sixty-three animals were assigned to treatment at 14 d before expected calving date and rema ined on the same treatment until 151 DIM. Four dietary treatments were the following: 1) no PHT prepartum or postpartum, 2) 1%

PAGE 25

12 PHT prepartum and 2% PHT postpartum, 3) 0% PHT prepartum and 2% PHT postpartum, and 4) 0% PHT prepartum and 2% PHT beginning at 35 DIM. Dry matter intake and milk production did not differ amon g treatments. When evaluating the first 35 d of lactation alone, cows fed fat beginning at calving had a 3.5 kg/d advantage of 3.5% FCM yield in comparison to cows fed fat beginning in the prepartum period, due in part to differences in the concentration of milk fat between the two treatments. Plasma NEFA concentrations, days to first estrus, pregna ncy rate, days open, and incidence of disease did not differ among treatments. Effects of Supplemental Fat on Adipose Tissue Lypolysis When the energy needed for maintenance a nd lactation is greater than the energy provided in the diet, the dairy cow will begin to mobilize her body fat stores to lessen the energy deficit. Nonesterified fatty acids are released into the bl ood by adipose tissue and transported to hepatic and non-hepatic tissues. Once in the liver, NEFA have the three following fates: 1) oxidized to carbon dioxide to provide energy, 2) partially oxidized for energy but also producing ke tone bodies such as -hydroxy butyric acid (BHBA) that serve as fuel for other tissues, or 3) reconve rted to triglycerides and stored (Drackley, 1999). When fat is added to the diet, plasma concentrations of NEFA routinely increase (Chilliard, 1993; Drackley, 1999; Grummer a nd Carroll, 1991). In a review of 50 treatment comparisons, Chilliard (1993) reported an average increase in concentration of plasma NEFA of 41 M (P < 0.005) over c ontrols when supplemental fat was fed. Likewise, Drackley (1999) reported an aver age increase in concentration of plasma NEFA of 81 M over controls when supplementa l fat was fed in reviewing seven studies. This increase due to dietary fat is much le ss than what is typica lly observed during the

PAGE 26

13 transition period when NEFA concentrati ons may increase up to 1 mM or more (Grummer, 1993). When NEFA concentrations in blood are high, as during the pe riparturient period, removal of NEFA by the liver may exceed the oxidative capacity of the liver and, when in combination with the low rate of export of triacylglycerol (TAG) out of the liver in ruminants, TAG accumulates in the liver (G rummer, 1993). Mechanisms to decrease hepatic lipid accumulation in cows fed fat might include decreased hepatic uptake of NEFA, increased oxidation of NEFA in the liver, decreased hepatic esterification of NEFA, or increased export of TAG from the liver (Grum et al., 1996). Illinois workers (Grum et al., 1996) conduc ted a study in which cows were fed a control diet (80% oat hay, DM basis), a high concentrate di et (51% oat hay, DM basis), or a high fat diet (6.5% of DM as EE) for 50 d prepartum and a common diet postpartum. Cows fed fat during the prepartum period tended to have decreased plasma NEFA concentrations early postpartum and had decr eased liver TAG concentrations at d 1 of lactation. They also reported a positive co rrelation between concen trations of plasma NEFA at 3 d prepartum and the concentration of TAG in the liver at 1 d postpartum. However, a similar study at Illinois (Douglas et al., 2004) in which cows were fed a moderate non-fiber carbohydrat e (NFC) control diet, a low NFC diet with 4% choice white grease (Qual-Fat) prepartum (DM basis), and a moderate NFC diet with 4% choice white grease (DM basis) beginning at 60 d prepar tum revealed no treatment differences in DMI, milk production, plasma concentration of NEFA, or total hepatic lipid or TAG at 1 DIM.

PAGE 27

14 During the periparturient period, the adipose tissue transfers from an anabolic state to a catabolic state. Tissue mobilization increases because the nutrient demands of the fetus and placenta are high and DMI dec lines during the last weeks of pregnancy (Grummer, 1993). Endocrine secretions ha ve a major impact on lipogenesis and lipolysis. Insulin increases glucose uptake by the cell membrane and increases lipogenic enzymes to stimulate FA and TAG synthesis. Staples et al. (1998) reviewed 17 studies that reported insulin concentr ations from fat-supplemented co ws and found mixed results. In 8 studies, concentration of plasma insulin of cows fed fa t were significantly depressed in comparison to controls, however differences were eliminated when adjusted for energy status of the animals. Leptin is a hormone synthesized by adipos e tissue that is pos itively regulated by adiposity and negatively by undernutrition. High concentrations of plasma leptin are associated with decreased feed intake and increased energy expenditure. Low concentrations of plasma le ptin are associated with in creased appetite and energy conservation. Concentrations of leptin ar e decreased around the time of parturition in concurrence with negative energy balance and a reduction in adipose stores and may be mediated by the reduction in plasma insu lin (Block et al., 2003). Circulating concentrations of plasma leptin were not different between beef heifers fed a basal diet or diets with supplemental fat (4% corn o il or 2% Ca salts of conjugated LA ( CLA ), DM basis) for 32 or 60 d before sl aughter (Gillis et al ., 2004). However, leptin concentrations in adipose tissue were greater for heifers s upplemented with 4% corn oil versus heifers fed the basal diet or 2% Ca salts of CLA (0.28, 0.18, and 0.17 g/g protein, respectively). Likewise, concen tration of plasma leptin of late lactation dairy cattle

PAGE 28

15 abomasally infused with cis-9, trans-11 or trans-10, cis-12 CLA isomers were not different (Baumgard et al., 2002). Effects of Supplemental Fa t on Immune Function In response to an activated immune syst em, the liver will produce acute phase proteins such as ceruloplasmin, fibrinoge n, and haptoglobin (Baumann and Gauldie, 1994). Haptoglobin is responsible for preventing the loss of body iron and concentrations are normally undetectable in bovi ne blood unless there is tiss ue damage. Ceruloplasmin is involved with copper transport and concentr ations will increase due to an inflammatory response of the cow. Fibrinogen is involved with blood clotting and the formation of the fibrin matrix for tissue repair. Increased fi brinogen concentrations are detected during internal hemorrhage or tissue damage. Arth ington and coworkers (2003) looked at the response of newly weaned beef calves to th e stresses of transpor tation and co-mingling and reported increased concentrations of plasma fibrinogen, ceruloplasmin, and haptoglobin. Immune reactions have been shown to be modulated by the diet, including the PUFA composition of the diet (Calder et al., 2002). Mechan isms involved in regulation are not yet understood, but evidence exists that PUFA composition of the diet influences cellular communication and activ ation through the synthesis of prostaglandins, tumor necrosis factor, and interferon(Calder et al., 2002). Linole ic acid can be converted to arachidonic acid, the precur sor for prostaglandin E2 and leukotriene B4 which are proinflammatory mediators. Similarly, LNA can be converted to eicosapentaenoic acid, the precursor for the synthesis of the in flammatory mediators prostaglandin E3 and leukotriene B5. Lessard et al. (2004) evaluated cellu lar immune functions of dairy cows fed supplemental fat during the transition peri od. Cows were fed diets of 2.7% Ca salts

PAGE 29

16 of palm oil (Megalac), 5.9% flaxseed (n -3 FA) or 9.4% micronized soybeans (n-6 FA) from 6 wk prepartum to calving. From calving to 6 wk postpartum, cows were fed diets of 4.7% Megalac, 9.7% flaxseed, or 20.3% micronized soybeans. Serum antibody response to ovalbumin injections during th e prepartum period did not differ among treatments. The lymphocyte response of blood mononuclear cells to mitogenic stimulation was lower in cows fed soybeans than in those receiving flaxseed or Megalac. The authors concluded that cellular immune functions were modulated around parturition; however feeding diets rich in n3 or n-6 FA did not have a major impact on cellular immune function. Immediately postpartum up to 90% of cows develop mild endometritis (Lewis, 1997). It is suggested by many research ers that a compromised immune system involving reduced effectiveness of neutroph ilic movement and pathogen destruction may be the reason that some cows spontaneously recover while others develop severe uterine infections that redu ce fertility (Lewis, 1997). Evalua tion of cervical discharge using vaginoscopy can be used as a diagnosis tool of uterine bacterial in fection (Dohmen et al., 1995). Clinical endometritis as described by a purulent or foul discharge after 20 d postpartum or a mucopurulent discharge afte r 26 d postpartum was associated with a reduction of pregnancy rates (LeBlanc et al., 2002). Abnormal vaginal discharge has been correlated with a delay in the firs t postpartum ovulation (O psomer et al., 2000). Furthermore, if first ovulation occurs in the presence of a uterus with heavy contamination, it can lead to prolonged luteal phases which is also associated with lower fertility (Opsomer et al., 2000).

PAGE 30

17 Effects of Supplemental Fat on Reproduction A Historical Review It is well established in nonruminants that animals with essential FA deficiencies have poor skin and hair, low growth rates, and reduced reproductive performance. In early studies, the essentiality of LA wa s documented primarily in nonruminants by causing then curing symptoms of deficiency. In the la te 1920s Burr and Burr (1929) established that dietary fat was essential to the growing rat. Afte r 70 d on purified diets, rats without lipid in the diet experienced dandruff, hair loss, cessation of growth, abnormal kidneys, blood in the urine, prol apsed penis, and irregular ovulation. All animals died after 120 to 230 d without diet ary fat. The following year, Burr and Burr (1930) sought to identify specific FA res ponsible for normal reproductive function in rats. Growing rats were fed lipid-free diets and upon weight loss, i ndividual lipid sources were supplemented at 1% of the diet. Of 22 females fed fat-free diets, 13 were not cycling or cycling irregularly. When three ovulatory rats that had been fed fat-free diets were bred, two produced litters; however no young lived more than a few hours. When four of the nonovulatory females were given five drops of either co rn (41% LA), olive (7% LA), linseed (59% LA), or coconut (1 % LA) oil daily, all re sumed ovulation except for the rat supplemented with coconut oil. Six of the i rregularly ovulating females fed fat-free diets were fed two drops of cod liver oil daily. After 4 wk all rats were cycling normally, were bred and produced normal litters. In response to their results, the authors stated, the resumption of ovulation is so rapid that growth has hardly begun. Synthesis of ovarian hormone ceases when fatty acids are eliminated from the diet. It was many years later that FA deficiency wa s studied in farm animals. In 1954 it was determined that the preweaned calf also requi res FA (Lambert et al., 1954). Preruminant

PAGE 31

18 calves were fed isocaloric, purif ied diets of synthetic milk de void of lipid with or without hydrogenated soybean oil and lecithin. Ca lves fed diets without lipid developed symptoms of deficiency including scaly dandr uff, long dry hair, hair loss, diarrhea, and low weight gain. Additionally, guinea pigs fed a basal fat-free diet for eight months had an average weight of 254 28 g, incidence of dermatitis was 75% and mortality rate was 25% while guinea pigs raised on the same diet and supplemented with 0.4% methyl linoleate (1.31% of calories) had an averag e weight of 382 15 g and had no dermatitis or mortality (Ried et al., 1964). Lastly, growing male pigs were fed 0, 0.25, 0.5, 1.0, 2.0, and 4.0% of dietary calories as LA using corn oil for ten wk (Sewell and McDowell, 1966). Concentration of LA in scrotal fat refl ected dietary intake after five and ten wk on diet. Skin lesions appeared on pigs recei ving 0, 0.25, and 0.5% of dietary calories as LA and they were then supplemented with met hyl linoleate or methyl oleate at 1.0% of dietary calories. Skin lesion disappeared in pigs supplemente d with methyl linoleate and remained on those supplemented with methyl oleate. It has been documented in nonruminants including the rat (Holtman, 1960), the guinea pig (Ried et al., 1964), and the pig (S ewell and McDowell, 1966) that a ratio of C20:3 to C20:4 in tissues/serum that exceeds 0.4 is indicative of a LA deficiency. The rational behind this ratio as an indicator of LA deficiency is that th e synthesis of C20:3 n9 from oleic acid increases when LA is de ficient because of enzyme competition. The Modern Dairy Cow Quantification of LA available for use by th e adult ruminant is difficult to predict because of ruminal biohydrogenation. Th e extent of ruminal biohydrogenation is variable and dependent upon many factors in cluding the diet and ruminal conditions, however it is estimated by Chilliard et al (2000) that 80% of dietary LA is

PAGE 32

19 biohydrogenated. Feeding ruminally inert fats, such as CSFA, will partially protect FA from biohydrogenation, allowing greater es cape to the small intestine. Using the fat sub-model of the CornellPenn-Miner (CPM)-Dairy model, Sanchez and Block (2002) suggested th at the amount of LA excret ed in 45.5 kg of milk daily exceeds the post ruminal uptake from typical di ets. Calculation of LA balance of the lactating dairy cow would be the following: LA absorbed from the diet LA used for maintenance LA used for milk production. Using the fat sub-model of the CPM-Dairy model, a cow consuming 25 kg of DM/d of a typical diet (no adde d fat source) would consume 225 g/d LA. Of the 225 g consume d, only 20% will es cape biohydrogenation (45 g), and of that, 82% (37 g) will be absorbed in the small intestine. The LA requirement for maintenance of the mature lactating ruminant has not been defined. However, a calculation based on metabolic body weight using the nonlactating rat (Mattos and Palmquist, 1977) yields a main tenance requirement of 10.7 g/d for a cow weighing 607 kg. Milk output of LA of a cow producing 45 kg milk/d would be 54 g/d (3.4% milk fat containing 3.5% LA). In this situation, the LA bala nce would be -27.7 g/d (37 g/d absorbed from the diet 10.7 g/d for maintenance 54 g/d for milk production). To get this animal out of a deficient situati on, a fat rich in LA must be supplemented. A possible explanation as to w hy numerous studies report an improvement in reproduction when additional fat is fed may be due to alle viating a LA deficiency of the modern highproducing dairy cow. Many studies report an improvement in reproductive performance of cows fed supplemental fat. In a review, Staples et al (1998) reported an improvement in fertility rates in 11 of 20 articles and speculated that it was a result of dietary FA and not solely

PAGE 33

20 due to an improvement in the energy st atus of the cows. Although reproductive performance is strongly associated with energy status (Staples et al., 1990), dietary fats can provide FA precursors for steroid (including cholesterol) and eicosanoid (including prostaglandins) production which have an a ffect on ovarian function, uterine function, and conception rates. Dietary fats typically increase concentra tions of circulating cholesterol, the precursor of progesterone (Grummer and Ca rroll, 1991). Ruminants fed supplemental fat often have a slight increase in blood progesterone concentrat ion (Staples et al., 1998). Progesterone, secreted by the corpus luteum ( CL ), prepares the uterus for implantation of the embryo and helps maintain pregnancy by providing nourishment for the conceptus via induction of heterotrophic pr oteins from the endometrium. Work by Hawkins et al. (1995) suggests that the increase seen in circulating progesterone when cows are fed supplemental fat is from a reduced rate of clearance of progester one rather than an increase in progesterone synthe sis. Son et al. (1996) repor ted greater blood cholesterol and peak plasma progesterone concentration during the second ovulatory cycle in cows fed tallow at 2 vs. 0% of dietary DM. Work ers at the University of Florida (GarciaBojalil et al., 1998) reported that accumulated plasma pr ogesterone from 0 to 50 DIM was greater, pregnancy rates im proved, and energy status did not change when cows were fed diets of 2.2% CSFA compared to non fat-supplemented cows. Through a series of desaturases and el ongases, LA (C18:2) can form dihomolinolenic acid, a direct precu rsor to the series 1 prosta glandins, or can be further desaturated to arachidonic acid (C 20:4), a direct precursor to the 2 series prostaglandins. Prostaglandin F2 ( PGF2 ), synthesized by endometrial tissu e, is an important regulator

PAGE 34

21 of parturition and the estrous cycle by causi ng regression of the CL. Upon conception, it is important to keep the CL from regressing in order to prevent early embryonic death. Immediately postpartum, 13, 14-dihydro-15-keto-PGF2 metabolite ( PGFM ) is important in regressing the CL of pregnancy. If LA is supplemented in the diet prepartum, more arachidonic acid may be synthesi zed leading to higher concen trations of the series 2 prostaglandins and possibly a healthier uterine environment. Alternately, if excess LA is consumed, it can be converted to eicosadienoi c acid (C20:2) instead of arachidonic acid (Kanduce et al., 1982), increasing the synthesis of the series 3 prostaglandins at the expense of the series 1 and 2 prostaglandins It is thought that LA can compete with arachidonic acid for binding si tes of a key enzyme, cyclooxygenase 2 (PGHS-2), that is necessary for the synthesis of PGF2 The amount of a particular FA (e.g. LA) stored in the target tissue may control whether there is an inhibition or stimul ation of prostaglandin synthesis. Reducing PGF2 secretion through dietary fats could improve pregnancy rates by reducing early embryonic loss aro und the time of embryo recognition. Plasma IGF-I concentrations are correl ated positively with body condition and DMI. Low IGF-I concentrations are associated with an extended postpartum interval to estrus in beef cows and also with delaye d puberty (Roberts et al ., 1997; Rutter et al., 1989), indicating that IGF-I can be positively correlated with reproductive performance. Beam and Butler (1998) reported lower mean c oncentrations of plasma IGF-I from wk 1 to 3 postpartum in cows fed a diet containi ng 2.6% prilled fat compared to controls (37.6 vs. 47.7 ng/ml) despite no differences in energy ba lance. However, other studies reported no differences in concentration of plasma IGF-I when supplemental fat was fed (Salado et al., 2004; Spicer et al., 1993) Insulin-like growth factor I acts synergis tically with

PAGE 35

22 luteinizing hormone ( LH ) to promote follicular development (Lucy, 2001). However, if IGF-I is over stimulated there may be dele terious effects on embryo development, the uterine environment, and gene expression (Bilby et al., 2004). More specifically, overstimulation of IGF-I is detrimental to fo llicle and oocyte development (Armstrong et al., 2001). Although the results are somewhat mixed, improvement in conception rates when fat is supplemented in the diet is often reported. In a study conducted in Wisconsin (Scott et al., 1995), five herds (n = 443) we re fed CSFA at 0 or 450 g/d from 1 to180 or 200 DIM. They reported an increase in ove rall conception rate from 93 to 98%, and a tendency for more fat-supplemented cows to exhibit standing estrus (71.4 vs. 65.6%). In addition, they reported a tendency for less incide nce of noncyclic ovaries in fat-fed cows. A study conducted in Pennsylvania and Israel by Ferguson et al. (1990) reported an improvement in first service conception rate when 253 cows over four herds were fed 0 or 2% ruminally inert fat from 0 to150 DIM (43 vs. 59%). Multiparous Holstein cows (n = 81) were fed isoenergetic diets containi ng 1.7% supplemental fat (prilled long chain FA) for 21 d prepartum and control or glucogenic-supplemented diets for 28 d postpartum (Frajblat and Butler, 2003). Fa t supplementation prepartum did not affect follicle dynamics measured by ultrasonagraphy nor concentration of plasma progesterone, insulin, IGF-I, or NEFA. However, supplemental fat prepartum was associated with better pregnancy rates (86 vs 58% for fat-supplemented and control cows respectively, P = 0.03). Recently, workers in Missouri (Oelrichs et al., 2004) reported no benefit for conception rates of Holstein cows (n = 64) fed raw, cracked soybeans beginning at 28 d prepartum or beginning at calving (fed at 1.9 and 2.9 kg of DM during

PAGE 36

23 the prepartum and postpartum periods, respec tively) despite an improvement in energy balance. Concentrations of plasma progest erone and PGFM, interval to first estrous cycle, and rates of cyclicity, ovulation, con ception and pregnancy we re not different from cows not fed soybeans. However, cows fed soybeans beginning either prepartum or at calving had fewer small (< 5 mm ) follicles and tended to have more medium (6 to 9 mm) follicles than controls during the first synchr onized estrous cycle. The high LNA and LA content in flaxseeds (57% LNA and 14% LA) may have been responsible for the improvement in conception rates (87.5 vs. 50.0%) of lactating dairy cows fed formaldehyde-treated whole flaxseed (17% of dietary DM) compared to those fed Ca salts of palm oil (5.6% of dietary DM) from 9 to 19 wk postpartum (Petit et al., 2001). To investigate the theory that specific FA (e.g., LA) reaching target tissues could improve conception rates, Santos and cowork ers (2004) supplemented dairy cows with a ruminally inert blend of LA and monoenoic trans FA or a Ca salt of palm oil from 25 d prepartum through ~55 d postpartum when cows were timed AI, then flushed 5 d after AI and recovered structures were evalua ted. Cows fed the LA and monoenoic trans FA tended to have (P = 0.11) a greater fertiliza tion rate (87 vs. 73%), had more accessory sperm per structure collected (34 vs. 21), and tended to have (P = 0.06) a greater proportion of embryos classified as high qua lity (73 vs. 51%). In an accompanying study, conception rate at firs t AI was greater for cows fed the blend of LA and monoenoic trans FA salt (38.9 vs. 25.9%). In contrast, a few studies ha ve reported a significant decr ease in conception rates of cows fed supplemental fat. In reviewing th ree studies that reporte d decreased conception rates, Staples et al. (1998) noted that in al l studies there was a dramatic improvement in

PAGE 37

24 milk production. High milk production and nega tive energy balance have been linked to decreased fertility in dairy cattle. Effects of Supplemental Fat on Follicle Development In addition to an improvement in concepti on rates of lactating dairy cows, follicular development is improved often by fat feeding. Cows in negative energy balance or in poor body condition can experience reduced ovari an activity (Staples et al., 1990) which might be alleviated quicker by supplementing with fats. The mechanism by which ovarian activity is affected by energy status is likely at the hypothalamuspituitary axis and perhaps at the ovary itself. Leutiniz ing hormone and follicle stimulating hormone ( FSH ) are secreted by the anterior pituit ary gland upon stimulation by gonadotropin releasing hormone ( GnRH ) released from the hypothalamus to cause recruitment and growth of ovarian follicles. In early l actation and during the st ate of negative energy balance, ovarian activity is reduced by low pulsatile secretion of LH (Beam and Butler, 1999). Simmetal cows (n = 12) were assigned to receive CSFA (0.5% of body weight) or an isocaloric control supplement in addition to prairie hay from parturition until the second postpartum ovulation. Calves were pe rmanently removed at 25 d postpartum to assist with a quicker return to estrus. Concentrations of mean serum LH and total cholesterol for fat-supplemented cows was great er than for control an imals. In addition, follicular development as determined by ultras onography was affected in that growth of class 2 (6 to 9 mm) follicles into class 3 (10 to 15 mm) and 4 (> 15 mm) follicles was enhanced in cows receiving CSFA (Hightshoe et al., 1991). At parturition, 18 cows were assigned to receive CSFA at 0 or 2.2% of dietary DM in a TMR containing 14.5% whole cottonseeds until 60 d postpartum (Lucy et al., 1991).

PAGE 38

25 Prior to 25 d postpartum, CSFA-supplemented cows had a decreased number of 3 to 5 mm follicles and an increased number of 6 to 9 mm follicles. After d 25 postpartum, estrous was synchronized. The number of 3 to 5 mm follicles and follicles > 25 mm increased in CSFA-fed cows. In addition, th e diameters of the larg est and second largest follicles were greater in CSFA-supplemented cows. Eighteen lactating Holstein cows were fe d CSFA at 2.2% of dietary DM or an isoenergetic diet (Lucy et al., 1993). Although animals were in similar energy balance, cows fed CSFA had a larger second wave do minant follicle (18.7 mm) than did cows fed the 0% CSFA diet (16.1 mm). Forty-five Holstein cows were fed a blend of tallow and yellow grease (88:12 wt/wt) at 0, 2.2, or 4.4% of the dietary DM fr om d 0 to 84 DIM (Beam and Butler, 1997). On d 14 postpartum, the number of follicl es greater than 15 mm in diameter was dramatically increased in cows fed diets of 2.2 and 4.4% (~ 0.7) supplemental fat in comparison to the control (~ 0.3) and was not correlated with energy status. The diameter of the largest follicle from d 8 to 14 postpartum was greater in cows fed 2.2% supplemental fat (13.5 mm) versus controls ( 11.0 mm). If only the animals that ovulated their first wave dominant follicle were considered, all fat-supplemented cows increased the diameter of the largest follicle from d 8 to 14 postpartum. At parturition, Holstein cows (n = 141) were allotted to one of three dietary treatments (Petit and Twagiramungu, 2002). The isonitrogenous, isoenergetic, and isolipidic diets contained whole flaxseed, Ca salts of palm oil, or micronized soybeans. The diameter of the CL of the cows fed flaxseed was larger than that of cows fed soybeans (19.7 vs.16.9 mm) but not larger than that of cows fed Ca salts of palm oil (17.5

PAGE 39

26 mm). Embryo mortality from day 30 to 50 af ter AI tended to be lower (P < 0.11) when cows were fed flaxseed (0%) compared to Ca salts of palm oil (15.4%) or soybeans (13.6%). The timing of initiation of fat supplementa tion during the periparturient period has received little attention. The beneficial results seen in vari ous studies when feeding fat is initiated during the periparturient period is not consistent. There is only one published study (Salfer et al., 1995) conducted to eval uate the timing of initiation of fat supplementation in lactating dairy cows. The authors concluded that delaying the inclusion of partially hydrogenated tallow in the diet until 35 DIM had benefits on total milk production through improved persistency. The objective of the current experiment was to evaluate if initiating fat suppl ementation during the prepartum period, at parturition, or at 28 DIM would have a be neficial effect on milk production, liver function, and reproduction of Ho lstein cows during summer.

PAGE 40

27Table 2.1. Effects of feeding ruminally inert fat (Ca salts of fatty acids (CSFA) or pril led fat (PF)) to the same cows through out the study on DMI, milk production, milk composition, and body weight (BW) change. Reference Treatments/ Fat source Dietary EE, % DIM Diet, % of DM DMI, kg/d Milk, kg/d 4.0% FCM, kg/d Milk protein, % Milk fat, % BW change, kg or kg/d Erickson et al., 1992 1) Control 2) 12 g/d niacin (NA) 3) 3% CSFA 4) NA + CSFA ------15-98 35% alfalfa haylage, 10% corn silage, 55% concentrate 1) 18.5 2) 18.5 3) 17.5 4) 18.2 1) 36.2 2) 36.4 3) 38.2 4) 39.3 F** 1) 32.4 2) 32.7 3) 34.3 4) 35.4 F** 1) 2.71 2) 2.84 3) 2.55 4) 2.68 F** 1) 3.32 2) 3.32 3) 3.36 4) 3.35 1) 0.18 2) 0.11 3) -0.22 4) 0.07 (kg/d) Moallem et al., 2000 1) Control 2) 0.55 kg/d CSFA 3) Control + bST ------0-150 9.5% wheat silage, 15% corn silage, 3.5% legume hay, 3.5% oat hay, 2.3% wheat straw, 66.2% concentrate 1) 23.5 2) 23.7 3) 24.5 1) 40.2 2) 42.4 3) 45.4 F** 1) 37.8 2) 40.8 3) 43.2 F** 1) 2.92 2) 2.92 3) 2.94 1) 3.12 2) 3.25 3) 3.19 F** N.S. Firkins and Eastridge, 1992 1) Control 2) 7% soy hulls (SH) 3) 7% SH + 1% NaHCO3 4) 20% SH + 0.43% CSFA + 10.7% roasted soybeans 1) 2.39 2) 2.39 3) 2.34 4) 3.42 (FA) 28-133 Trt 1 and 4: 10% alfalfa silage, 31% corn silage, 59% concentrate Trt 2 and 3: 10% alfalfa silage, 20% corn silage, 70% concentrate 1) 24.4 2) 23.5 3) 23.2 4) 22.5 1) 37.0 2) 37.3 3) 35.1 4) 38.9 1) 33.8 2) 33.0 3) 31.2 4) 35.6 2 & 3 vs. 4* 1) 3.16 2) 3.18 3) 3.18 4) 2.88 F* 1) 3.50 2) 3.26 3) 3.39 4) 3.42 2 & 3 vs. 4* 1) 0.38 2) 0.43 3) 0.28 4) 0.18 (kg/d) Garcia-Bojalil et al., 1998 1) 11.1% RDP 2) 11.1% RDP + 2.2% CSFA 3) 15.7% RDP 4) 15.7% RDP + 2.2% CSFA 1) 4.77 2) 6.65 3) 4.62 4) 6.20 0-120 34% corn silage, 13% alfalfa hay, 53 % concentrate a 1) 19.6 2) 19.0 3) 19.4 4) 19.8 F x RDP ** at 0-50 DIM 1) 27.1 2) 28.0 3) 25.5 4) 27.7 F** at 50120 DIM 1) 25.5 2) 26.5 3) 24.1 4) 26.3 1) 3.06 2) 3.02 3) 3.06 4) 2.98 F** at 35120 DIM 1) 3.63 2) 3.67 3) 3.66 4) 3.68 N.S. Spicer et al., 1993 1) Control 2) 1.8% CSFA ------28-84 20% sorghum silage, 19% alfalfa hay, 61% concentrate a 1) 25.9 2) 24.4 1) 36.9 2) 36.0 1) 34.1 2) 32.7 ------1) 3.54 2) 3.44 1) 1.4 2) 6.3 F** 3.5% FCM. a Concentrate mix included 6-15% whole cottonseed. F = effect of fat. N.S. = not significant. P < .10. ** P < .05.

PAGE 41

28Table 2.1. Continued. Reference Treatments/ Fat source Dietary EE, % DIM Diet, % of DM DMI, kg/d Milk, kg/d 4.0% FCM, kg/d Milk protein, % Milk fat, % BW change, kg or kg/d Chouinard et al., 1997 1) Control + NaHCO3 2) 2% CSFA + NaHCO3 3) 4% CSFA + NaHCO3 4) 4% CSFA 1) 3.9 2) 5.5 3) 6.6 4) 6.9 39-67 62% alfalfa silage, 38% concentrate 1) 23.6 2) 22.8 3) 21.4 4) 21.6 F** (linear) 1) 33.5 2) 35.2 3) 32.8 4) 32.0 F** (quadratic) 1) 31.2 2) 31.6 3) 28.9 4) 28.0 F** (linear) 1) 2.96 2) 2.80 3) 2.66 4) 2.57 F** 1) 3.55 2) 3.38 3) 3.29 4) 3.43 ------Chouinard et al., 1997 1) Control + NaHCO3 2) 2% CSFA + NaHCO3 3) 4% CSFA + NaHCO3 4) 4% CSFA 1) 4.0 2) 5.8 3) 7.5 4) 7.5 68-95 46% alfalfa silage, 54% concentrate 1) 23.4 2) 21.9 3) 21.5 4) 21.0 1) 33.6 2) 35.9 3) 34.0 4) 31.4 F x NaHCO3* 1) 30.4 2) 31.3 3) 28.7 4) 26.7 1) 3.10 2) 2.96 3) 2.82 4) 2.74 F** 1) 3.41 2) 3.08 3) 2.93 4) 3.23 F** linear ------Harrison et al., 1995 1) Control 2) 12% WCS 3) 12% WCS and 2.7% CSFA 1) 2.5 2) 4.4 3) 6.0 21-119 23% alfalfa hay, 23% grass silage, 54% concentrate 1) 23.1 2) 23.9 3) 21.6 Trt** ------1) 38.1 2) 39.8 3) 39.5 Trt** 1) 3.08 2) 3.07 3) 2.91 Trt** 1) 3.24 2) 3.49 3) 3.74 Trt** ------Harrison et al., 1995 1) Control 2) 12% WCS 3) 12% WCS and 5% CSFA 1) 3.5 2) 5.1 3) 6.9 18-105 28% alfalfa hay, 18% grass silage, 54% concentrate 1) 23.6 2) 22.4 3) 22.2 Trt** Trt x P** ------1) 36.0 2) 37.2 3) 37.8 Trt** Trt x P** 1) 3.09 2) 3.11 3) 2.95 Trt** Trt x P** 1) 3.36 2) 3.65 3) 3.72 Trt** Trt x P** ------Holter et al., 1992 1) Control 2) 15% WCS 3) 15% WCS + 0.54 kg/d CSFA ------0-112 Ad libitum forage: 63% corn silage, 37% wilted grass silage. Concentrate was adjusted to milk production 1) 17.4 2) 16.6 3) 16.8 1) 35.2 2) 29.6 3) 32.5 Trt** 1) 31.6 2) 30.3 3) 31.9 1) 2.86 2) 2.88 3) 2.82 Trt** 1) 3.32 2) 4.14 3) 3.89 Trt** ------3.5% FCM. WCS = whole cottonseed. F = effect of fat. Trt = effect of treatment. P = effect of parity. N.S. = not significant. P < .10. ** P < .05.

PAGE 42

29Table 2.1. Continued. Reference Treatments/ Fat source Dietary EE, % DIM Diet, % of DM DMI, kg/d Milk, kg/d 4.0% FCM, kg/d Milk protein, % Milk fat, % BW change, kg or kg/d Selberg et al., 2004 1) Control 2) 225 g/d CaS conjugated linoleic acid 3) 225 g/d CaS transC18:1 Prepartum control: 4.3 Postpartum control: 5.2 -28-49 Prepartum: 13% bermuda grass hay, 39% corn silage, 52% concentrate Postpartum: 10% alfalfa hay, 29% corn silage, 61% concentrate 1) 21.6 2) 20.0 3) 20.2 1 vs. 3 at wk 4-6** 1 vs. 2 at wk 6** 1) 40.3 2) 41.5 3) 41.5 1 & 2 vs. 3 by wk** 1) 40.0 2) 37.4 3) 40.5 1) 2.89 2) 2.82 3) 2.81 1) 3.49 2) 2.99 3) 3.46 1 vs. 2** ------Hoffman et al., 1991 1)Degradable protein (DP) 2) Undegradable protein (UP) 3) DP + 2.8% sodium alginate treated tallow (SAT) 4) UP + 2.8% SAT 1) 3.1 2) 3.3 3) 5.7 4) 6.0 22-150 49% alfalfa silage, 51% concentrate 1) 22.7 2) 22.5 3) 22.7 4) 22.7 (group fed) 1) 31.8 2) 31.6 3) 33.1 4) 32.7 F x T** 1) 29.9 2) 29.9 3) 30.9 4) 31.2 1) 3.14 2) 3.03 3) 3.04 4) 3.00 F** F x T** 1) 3.67 2) 3.63 3) 3.60 4) 3.82 N.S. Beam and Butler, 1998 1) Control 2) 2.59% PF 1) 4.8 2) 7.0 0-100 26% corn silage, 18% alfalfa haylage, 56% concentratea 0-100 DIM N.S. 0-28 DIM 1) 15.5 2) 17.3 F** F x T* (Control peaked ~41 vs. 43 kg/d for PF) F x T* (Control peaked ~34 vs. 36 kg/d for PF) ------------1) -26.5 2) -42.6 F* Jerred et al., 1990 b 1) Low silage (LS) 2) LS + 5% PF 3) Medium silage (MS) 4) MS + 5% PF 5) High silage (HS) 6) HS + 5% PF 1) 3.1 2) 6.5 3) 3.4 4) 7.2 5) 3.8 6) 7.2 5-105 Trt 1 & 2: 45% alfalfa silage, 55% concentrate Trt 3 & 4: 64% alfalfa silage, 36% concentrate Trt 5 & 6; 84% alfalfa silage, 16% concentrate 1) 23.6 2) 22.1 F** F x T** 1) 39.2 2) 38.8 1) 36.5 2) 37.8 F x T** 1) 2.89 2) 2.87 F x T** 1) 3.57 2) 3.88 F** 1) -0.36 2) -0.47 (kg/d) 3.5% FCM. a Concentrate mix included 6-15% whole cottonseed. b Note: there are no fat x forage interactions ; results are presented as 1) control (t reatments 1, 3 and 5) and 2) fat (treatme nts 2, 4, and 6). F = effect of fat. T = effect of time. N.S. = not significant. P < .10. ** P < .05.

PAGE 43

30Table 2.1. Continued. Reference Treatments/ Fat source Dietary EE, % DIM Diet, % of DM DMI, kg/d Milk, kg/d 4.0% FCM, kg/d Milk protein, % Milk fat, % BW change, kg or kg/d Skaar et al., 1989 1) Control 2) 12 g/d niacin (NA) 3) 5% PF 4) 12 g/d NA and 5% PF 1) 3.4 b 2) 3.4 b 3) 11.8 b 4) 11.8 b -17-105 25% corn silage, 25% alfalfa silage, 50% concentrate 1) 19.3 2) 17.8 3) 19.2 4) 18.7 1) 38.4 2) 36.3 3) 42.0 4) 41.3 F x season** (fat greater in summer) 1) 36.3 2) 34.5 3) 39.3 4) 38.2 F x season** (fat was greater in summer) 1) 3.00 2) 2.87 3) 2.87 4) 2.87 1) 3.14 2) 3.19 3) 3.15 4) 3.12 F x T** (fat fed gained faster) Pickett et al., 2003 1) Control 2) 500 ml/d propylene glycol (PG) drench 3) 0.45 kg/d CSFA drench 4) 500 ml/d PG + 0.45 kg/d CSFA drench 4.8 0-21 31% corn silage, 16% alfalfa hay, 9% alfalfa hay, 44% concentrate a Drenches were administered from 0-3 DIM 1) 17.2 2) 18.0 3) 16.9 4) 15.8 1) 36.5 2) 36.1 3) 32.5 4) 34.8 1) 42.9 2) 40.3 3) 38.1 4) 40.9 1) 3.68 2) 3.46 3) 3.66 4) 3.51 1) 4.66 2) 4.30 3) 4.64 4) 4.66 ------Schroeder et al., 2003 1) TMR fed (control) 2) Pasture + 6.7 kg/d corn based concentrate 3) Pasture + 6.7 kg/d concentrate with 0.8 kg CSFA 1) 4.5 2) 6.1 3) 8.1 117-152 Control: 59% corn silage, 41% concentrate Treatments 2 and 3: 84% pasture, 16% concentrate 1) 23.7 2) 22.9 3) 21.5 1) 20.2 2) 19.2 3) 20.2 1) 19.5 2) 17.8 3) 16.1 1 vs. 3** 1) 3.70 2) 3.49 3) 3.41 1 vs. 2 & 3** Trt x T** 1) 3.91 2) 3.45 3) 2.56 Trt** Trt x T** 1) 23 2) -6 3) -10 1 vs. 2 & 3** Moallem et al., 1999 1) Control 2) 0.55 kg/d CSFA 3) Control + bST ------0-150 8% wheat silage, 20% corn silage, 3% pea hay, 3% oat hay, 66% concentrate a 1) 24.0 2) 23.3 3) 24.7 (group fed) 1) 39.7 2) 42.5 3) 44.0 F** 1) 37.8 2) 40.9 3) 41.9 F** 1) 2.98 2) 2.92 3) 2.92 F** 1) 3.18 2) 3.25 3) 3.19 Fat fed lost more BW (~7 kg) than control (P < 0.1) Kim et al., 1993 1) Control 2) 17% extruded soybeans 3) 4.0% CSFA 1) 2.5 2) 5.1 3) 3.0 28-105 25% alfalfa hay, 25% corn silage, 50% concentrate 1) 17.8 2) 18.4 3) 16.6 2 vs. 3* 1) 29.2 2) 32.4 3) 31.8 F** 1) 25.4 2) 26.7 3) 28.7 F* 1) 2.99 2) 2.93 3) 2.81 F** 2 vs. 3** 1) 3.20 2) 2.69 3) 3.47 2 vs. 3** ------3.5% FCM. a Concentrate mix included 6-15% whole cottonseed. b concentrate mix only. F = effect of fat. T = effect of time. Trt = effect of treatment. N.S. = not significant. P < .10. ** P < .05.

PAGE 44

31Table 2.1. Continued. Reference Treatments/ Fat source Dietary EE, % DIM Diet, % of DM DMI, kg/d Milk, kg/d 4.0% FCM, kg/d Milk protein, % Milk fat, % BW change, kg or kg/d Moallem et al., 1997 1) Control 2) 0.5 kg/d CSFA 3) Control + bST 4) 0.5 kg/d CSFA + bST ------0-150 10.4% wheat silage, 24% corn silage, 2.3% pea hay, 2.1% oat hay, 61.2% concentrate 1) 24.1 2) 24.3 3) 24.9 4) 24.7 (group fed) 1) 36.3 2) 39.8 3) 41.3 4) 42.9 F** F x bST** 1) 33.4 2) 36.9 3) 39.5 4) 40.8 F** F x bST** 1) 3.02 2) 2.99 3) 3.01 4) 2.99 1) 2.98 2) 3.03 3) 3.21 4) 3.20 1) 38.0 2) 43.2 3) 33.4 4) 35.4 (maximum BW loss) Sklan et al., 1994 1) Control 2) 2.5% CSFA 1) 2.8 2) 4.9 0-120 14.3% wheat silage, 15.3% corn silage, 6.4% vetch hay, 64% concentrate a 1) 20.6 2) 20.3 (group fed) 1) 31.1 2) 35.0 F** F x T** 1) 29.4 2) 33.8 F** F x T** 1) 2.97 2) 2.99 F x T** 1) 3.15 2) 3.26 F* F x T** N.S. Cervantes et al., 1996 1) Control 2) 0.4 kg/d CSFA 3) 12 g/d nicotinamide (NM) 4) 0.4 kg/d CSFA + 12 g/d NM 1) 3.1 2) 5.1 3) 3.0 4) 5.0 112-150 Varied dependent on stage of lactation. Forages (35-60% of diet) utilized were alfalfa hay, alfalfa haylage, and corn silage. 1) 20.3 2) 20.5 3) 24.0 4) 21.1 F x NM* 1) 30.7 2) 31.8 3) 33.5 4) 33.2 1) 28.3 2) 29.7 3) 29.7 4) 30.6 1) 3.21 2) 3.17 3) 3.31 4) 3.14 F** 1) 3.45 2) 3.57 3) 3.26 4) 3.46 F* ------Palmquist and Weiss, 1994 1) Control 2) 2.5% tallow + 2.5% CSFA (Also included 3 concentrations of RUP) 1) 3.08 2) 6.73 (FA) 0-60 25% corn silage, 25% alfalfa hay, 50% concentrate 1) 18.1 2) 18.8 1) 43.3 2) 42.4 1) 37.1 2) 37.1 1) 2.97 2) 2.93 1) 2.64 2) 2.78 1) -0.08 2) -0.23 (kg/d) Atwal et al,, 1990 1) Control 2) 5% CSFA 1) 2.4 14-70 15% alfalfa silage, 10% alfalfa hay, 25% corn silage, 50% concentrate 1) 19.2 2) 19.7 1) 33.1 2) 33.3 1) 31.1 2) 32.5 1) 2.93 2) 2.94 1) 3.51 2) 3.80 1) 0.10 2) 1.35 (kg/d) F** (wk 1-4) 3.5% FCM. a Concentrate mix included 6-15% whole cottonseed. F = effect of fat. T = effect of time. P < .10. ** P < .05.

PAGE 45

32Table 2.1. Continued. Reference Treatments/ Fat source Dietary EE, % DIM Diet, % of DM DMI, kg/d Milk, kg/d 4.0% FCM, kg/d Milk protein, % Milk fat, % BW change, kg or kg/d Atwal et al,, 1990 1) Control 2) 5% CSFA 1) 2.3 1-56 25% alfalfa hay, 25% corn silage, 50% concentrate 1) 16.5 2) 15.1 1) 30.1 2) 29.7 1) 32.6 2) 30.8 1) 3.03 2) 3.01 1) 4.82 2) 4.37 1) -0.35 2) -1.85 (kg/d) Simas et al., 1995 1) Dry rolled sorghum (DRS) 2) Steam flaked sorghum (SFS) 3) DRS + 2.5% CSFA 4) SFS + 2.5% CSFA 1) 5.7 2) 5.7 3) 7.5 4) 7.5 5-91 34% alfalfa hay, 66% concentrate1 1) 21.6 2) 23.1 3) 17.8 4) 19.5 F** 1) 34.3 2) 39.3 3) 33.4 4) 36.5 1) 31.6 2) 34.5 3) 31.2 4) 32.8 1) 2.93 2) 3.00 3) 2.79 4) 2.99 3 vs. others* 1) 3.16 2) 2.91 3) 3.23 4) 3.05 1) 0 2) 0.16 3) -0.2 4) -0.02 (kg/d) F* Sklan et al., 1991 1) Control 2) 2.6% CSFA ------0-120 13.7% corn silage, 11.3% vetch hay, 75% concentrate 1 1) 20.3 2) 20.2 (group fed) 1) ~37 2) ~39 (at peak) F** at 30 and 60 DIM 1) ~34 2) ~38 (at peak) F** until 90 DIM N.S. 1) ~2.8 2) ~3.0 F** at 30 to 90 DIM F** (CSFA cows lost more and faster) Sklan et al., 1992 1) Control 2) 2% FA 3) 2.4% CSFA 1) 2.1 2) 4.2 3) 4.2 (FA) 93-213 1.4% oat hay, 20% wheat silage, 13.7% corn cobs, 64.9% concentrate 1) 21.2 2) 21.1 3) 20.9 (group fed) 1) 30.2b 2) 31.3a 3) 30.9ab Trt x T** Trt x P** 1) 25.9c 2) 27.7b 3) 27.0a Trt x T** Trt x P** 1) 3.20 2) 3.13 3) 3.15 Trt x T** Trt x P** 1) 2.67a 2) 2.81b 3) 2.75ab Trt x T** Trt x P** ------Sklan et al., 1992 1) 14.5% WCS 2) 1.8% FA 3) 2.1% CSFA 1) 5.0 2) 4.0 3) 4.0 (FA) 108-228 7.8% citrus silage, 32.7% wheat silage, 6.7% vetch hay, 52.8% concentrate 1) 20.1 2) 20.4 3) 20.3 (group fed) 1) 31.3b 2) 32.5a 3) 32.4ab Trt x T** 1) 30.1 2) 29.7 3) 30.4 Trt x T** 1) 3.02 2) 2.96 3) 2.98 Trt x T* 1) 3.25a 2) 2.99b 3) 3.17a ------3.5% FCM. a,b,c Means not followed by the same letter differ (P < .05). 1 Concentrate mix included 6-15% whole cottonseed (WCS). F = effect of fat. Trt = effect of treatment. T = effect of time. P = effect of parity. N.S. = not significant. P < .10. ** P < .05.

PAGE 46

33Table 2.2. Effects of feeding oilseeds alone or in combination with other fat sources to the same cows throughout the experimen t on DMI, milk production, milk composition, and body weight (BW) change. Reference Treatments/ Fat source Dietary EE, % DIM Diet, % of DM DMI, kg/d Milk, kg/d 4.0% FCM, kg/d Milk protein, % Milk fat, % BW change, kg or kg/d Stegeman et al., 1992 1) Control 2) Control + bST 3) 10% rolled sunflower seeds + bST 4) 10% rolled safflower seeds + bST 1) 2.3 2) 2.3 3) 6.2 4) 5.5 104-216 25% corn silage, 25% alfalfa hay, 50% concentrate 1) 27.5 2) 23.9 3) 27.3 4) 19.3 3 vs. 4** 1) 29.5 2) 32.7 3) 40.0 4) 34.1 2 vs. 3 & 4** 3 vs. 4** 1) 25.0 2) 27.9 3) 32.3 4) 28.1 3 vs. 4** 1) 3.17 2) 3.27 3) 3.02 4) 3.08 F** 2 vs. 3 & 4** 1) 2.99 2) 3.06 3) 2.73 4) 2.86 1) 24.7 2) 19.4 3) 36.1 4) 39.9 AbuGhazaleh et al., 2004 1) Control 2) 10.64% extruded soybeans + 5.5% fish meal 1) 2.67 2) 4.93 103-173 25% alfalfa hay, 25% corn silage, 50% concentrate 1) 29.3 2) 27.7 F* 1) 34.5 2) 38.9 F** F x T** 1) 36.0 2) 36.4 1) 3.39 2) 3.18 F** F x T** 1) 3.74 2) 3.17 F** F x T** ------Kim et al., 1993 1) Control 2) 17% extruded soybeans 3) 4.0% CSFA 1) 2.5 2) 5.1 3) 3.0 28-105 25% alfalfa hay, 25% corn silage, 50% concentrate 1) 17.8 2) 18.4 3) 16.6 2 vs. 3* 1) 29.2 2) 32.4 3) 31.8 F** 1) 25.4 2) 26.7 3) 28.7 F* 1) 2.99 2) 2.93 3) 2.81 F** 2 vs. 3** 1) 3.20 2) 2.69 3) 3.47 2 vs. 3** ------Markus et al., 1996 1) Control 2) 7.1% whole sunflower seeds 3) 2.7% tallow 1) 1.8 2) 4.2 3) 4.1 16-112 12% corn silage, 14% alfalfa silage, 9.5% alfalfa hay, 64.5% concentrate 1) 22.2 2) 21.1 3) 21.6 1) 34.4 2) 34.6 3) 35.5 1) 30.0 2) 29.9 3) 31.6 1) 3.1 2) 3.0 3) 3.0 1) 3.2 2) 3.1 3) 3.3 ------Weiss and Wyatt, 2003 1) Control 2) 12.3% whole roasted soybeans 3) 2.35% tallow (Also included 3 levels of vitamin E) ------160-188 38% corn silage, 8% alfalfa hay, 7% alfalfa silage, 47% concentrate 1) 22.3 2) 24.0 3) 22.0 2 vs. 3** 1) 35.1 2) 36.8 3) 37.5 F* ------1) 2.97 2) 2.92 3) 2.86 1) 3.76 2) 3.83 3) 3.08 F** 2 vs. 3** 1) 0.87 2) 1.29 3) 0.71 (kg/d) 2 vs. 3** 3.5% FCM. F = effect of fat. T = effect of time. P < .10. ** P < .05.

PAGE 47

34Table 2.2. Continued. Reference Treatments/ Fat source Dietary EE, % DIM Diet, % of DM DMI, kg/d Milk, kg/d 4.0% FCM, kg/d Milk protein, % Milk fat, % BW change, kg or kg/d Drackley et al., 1998 1) Control 2) Control + 12 g/d niacin 3) 10% whole raw soybeans and 2.5% tallow 4) 10% whole raw soybeans, 2.5% tallow and 12 g/d niacin 1) 2.75 2) 2.75 3) 6.04 4) 6.04 (FA) 28-301 32.5% alfalfa haylage, 17.5% corn silage, 50% concentrate 1, 2 1) 21.9 2) 21.7 3) 21.6 4) 22.2 F x T** 1) 30.5 2) 33.2 3) 31.8 4) 33.6 F x T** 1) 30.4 2) 32.9 3) 32.5 4) 34.2 F x T** 1) 3.29 2) 3.16 3) 3.16 4) 3.13 F** F x T** 1) 3.56 2) 3.50 3) 3.68 4) 3.60 F x niacin x T** Sklan et al., 1992 1) Control 2) Control + 2.4% FA3 3) Control + 16.2% WCS 1) 1.8 2) 4.4 3) 4.4 (FA) 70-210 8.4% alfalfa hay, 17.2% corn silage, 74.4% concentrate 1) 21.4 2) 22.1 3) 21.4 (group fed) 1) 32.7 2) 33.6 3) 33.5 Trt x T** 1) 27.6b 2) 28.7ab 3) 30.4a Trt x T ** 1) 2.97 2) 2.95 3) 2.96 Trt x T* 1) 2.54b 2) 2.57b 3) 2.96a Trt x T* ------Sklan et al., 1992 1) 14.5% WCS 2) 1.8% FA3 3) 2.1% CSFA 1) 5.0 2) 4.0 3) 4.0 (FA) 108-228 7.8% citrus silage, 32.7% wheat silage, 6.7% vetch hay, 52.8% concentrate 1) 20.1 2) 20.4 3) 20.3 (group fed) 1) 31.3b 2) 32.5a 3) 32.4ab Trt x T** 1) 30.1 2) 29.7 3) 30.4 Trt x T** 1) 3.02 2) 2.96 3) 2.98 Trt x T* 1) 3.25a 2) 2.99b 3) 3.17a ------1 Concentrate mix included 6-15% whole cottonseed (WCS). 2 After 175 DIM, diets were adjusted for decreased nutrient requireme nts. Forage content increased to 60% of DM. For treatments 3 and 4 whole raw soybeans were removed and tallow was decreased to 2.25% of the diet. 3 Fatty acid (FA) source was mixed soapstock contai ning 86% free FA, of which 20% was linoleic acid. a,b,c Means not followed by the same letter differ (P < .05). 3.5% FCM. F = effect of fat. Trt = effect of treatment. T = effect of time. P < .10 ** P < .05

PAGE 48

35Table 2.2. Continued. Reference Treatments/ Fat source Dietary EE, % DIM Diet, % of DM DMI, kg/d Milk, kg/d 4.0% FCM, kg/d Milk protein, % Milk fat, % BW change, kg or kg/d Schingoethe and Casper, 1991 1) Control 2) Added fat from extruded soybeans or sunflower seeds (summary of 5 studies) 1) 2.6 2) 5.3 (average) 28-119 38% corn silage, 12% alfalfa hay, 50% concentrate 1) 20.8 2) 20.4 1) 3000 2) 3085 (kg) ------1) 2.99 2) 2.91 F** 1) 3.20 2) 2.95 F** 1) 0.18 2) 0.13 (kg/d) Harrison et al., 1995 1) Control 2) 12% WCS 3) 12% WCS and 2.7% CSFA 1) 2.5 2) 4.4 3) 6.0 21-119 23% alfalfa hay, 23% grass silage, 54% concentrate 1) 23.1 2) 23.9 3) 21.6 Trt** ------1) 38.1 2) 39.8 3) 39.5 Trt** 1) 3.08 2) 3.07 3) 2.91 Trt** 1) 3.24 2) 3.49 3) 3.74 Trt** ------Harrison et al., 1995 1) Control 2) 12% WCS 3) 12% WCS and 5% CSFA 1) 3.5 2) 5.1 3) 6.9 18-105 28% alfalfa hay, 18% grass silage, 54% concentrate 1) 23.6 2) 22.4 3) 22.2 Trt** Trt x P** ------1) 36.0 2) 37.2 3) 37.8 Trt** Trt x P** 1) 3.09 2) 3.11 3) 2.95 Trt** Trt x P** 1) 3.36 2) 3.65 3) 3.72 Trt** Trt x P** ------Khorasani et al., 1991 1) Control 2) 4.5% jet-sploded whole canola seed (JSWCS) 3) 9% JSWCS 4) 13.2% JSWCS 5) 17.4% JSWCS 1) 2.2 2) 3.4 3) 4.4 4) 5.5 5) 6.7 36-92 30% alfalfa silage, 10% oat silage, 60% concentrate 1) 17.8 2) 18.8 3) 18.3 4) 16.2 5) 17.0 1) 32.5 2) 34.5 3) 34.0 4) 33.2 5) 29.8 1) 27.7 2) 28.2 3) 29.8 4) 26.9 5) 24.8 1) 2.95 2) 3.12 3) 2.89 4) 2.84 5) 2.71 F** (linear) 1) 3.08 2) 2.89 3) 3.06 4) 2.76 5) 2.84 ------Faldet and Satter, 1991 1) 10% soybean meal 2) 13% raw soybeans (RS) 3) 13% heated soybeans (HS) 1) 3.3 2) 5.6 3) 5.6 15-119 50% alfalfa silage, 50% concentrate 1) 23.4 2) 22.3 3) 23.6 1) 34.5 2) 34.2 3) 38.9 HS** 1) 33.4 2) 34.7 3) 38.0 HS** 1) 2.99 2) 2.89 3) 2.85 HS** 1) 3.41 2) 3.50 3) 3.41 1) 26 2) 2 3) 31 RS** 3.5% FCM. WCS = whole cottonseeds. F = effect of fat. P = effect of parity. Trt = effect of treatment. P < .10. ** P < .05.

PAGE 49

36Table 2.2. Continued. Reference Treatments/ Fat source Dietary EE, % DIM Diet, % of DM DMI, kg/d Milk, kg/d 4.0% FCM, kg/d Milk protein, % Milk fat, % BW change, kg or kg/d Holter et al., 1992 1) Control 2) 15% WCS 3) 15% WCS + 0.54 kg/d CSFA ------0-112 Ad libitum forage: 63% corn silage, 37% wilted grass silage. Concentrate was adjusted to milk production 1) 17.4 2) 16.6 3) 16.8 1) 35.2 2) 29.6 3) 32.5 Trt** 1) 31.6 2) 30.3 3) 31.9 1) 2.86 2) 2.88 3) 2.82 Trt** 1) 3.32 2) 4.14 3) 3.89 Trt** ------Wu et al., 1994 1) Control 2) 12% WCS 3) 12% WCS + 2.2% safflower oil 4) 12% WCS +2.2% prilled tallow 5) 12% WCS +4.4% prilled tallow 1) 3.3 2) 5.2 3) 7.4 4) 7.4 5) 9.6 50-125 43% alfalfa hay, 57% concentrate 1) 28.2 2) 27.2 3) 28.8 4) 26.8 5) 24.1 1) 32.5 2) 32.6 3) 35.0 4) 34.3 5) 33.0 1 & 2 vs. 3 &4** 1) 32.4 2) 32.3 3) 33.4 4) 34.6 5) 33.0 1 & 2 vs. 3 &4** 1) 3.20 2) 3.03 3) 3.03 4) 3.08 5) 3.07 1 vs. 2 ** 1) 3.49 2) 3.48 3) 3.26 4) 3.58 5) 3.51 1) 0.70 2) 0.82 3) 0.57 4) 0.64 5) 0.46 (kg/d) Kim et al., 1991 1) Basal containing 14% soybean meal (SBM) 2) 17% extruded soybeans (ESB) 3) 17% ESB and 5% SBM 1) 2.6 2) 5.5 3) 5.1 28-112 25% corn silage, 25% alfalfa hay, 50% concentrate 1) 20.9 2) 20.7 3) 19.8 1) 33.0 2) 35.8 3) 34.2 F** 1) 28.5 2) 29.6 3) 30.2 1) 2.92 2) 2.88 3) 2.83 1) 3.20 2) 2.88 3) 3.17 2 vs. 3* 1) 0.31 2) 0.00 3) -0.02 (kg/d) Pires et al., 1996 1) Control 2) 18% ground roasted soybean 3) 18% whole soybean 4) 2.7% blood meal 5) 2.7% blood meal +3% tallow 1) 3.2 2) 6.2 3) 6.2 4) 3.2 5) 6.2 21-126 30% corn silage, 20% alfalfa silage, 50% concentrate 1) 23.6a 2) 22.7ab 3) 21.3bc 4) 21.3bc 5) 20.4c 1) 39.6 2) 40.7 3) 36.4 4) 36.1 5) 39.3 1) 35.4 2) 35.0 3) 33.3 4) 33.9 5) 35.0 1) 3.03a 2) 2.83b 3) 2.88bc 4) 3.08a 5) 2.98ac 1) 3.33 2) 3.09 3) 3.50 4) 3.63 5) 3.29 ------a,b,c Means not followed by the same letter differ (P < .05). 3.5% FCM. WCS = whole cottonseeds. Trt = effect of treatment. P < .10. ** P < .05.

PAGE 50

37Table 2.3. Effects of feeding rendered fats to the same cows throughout the study on DMI, milk production, milk composition and body weight (BW) change. Reference Treatments/ Fat source Dietary EE, % DIM Diet, % of DM DMI, kg/d Milk, kg/d 4.0% FCM, kg/d Milk protein, % Milk fat, % BW change, kg or kg/d Tackett et al., 1996 1) 21% NDF 2) 21% NDF + 6% choice white grease (CWG) 3) 28% NDF 4) 28% NDF + 6% CWG 1) 2.8 2) 8.4 3) 2.7 4) 7.8 32-122 Treatment 1 & 2: 36% corn silage, 9% alfalfa hay, 55% concentrate Treatment 3 & 4: 36% corn silage, 16% alfalfa hay, 48% concentrate 1) 24.4 2) 21.7 3) 23.7 4) 21.5 1) 37.5 2) 38.9 3) 34.7 4) 38.0 1) 37.0 2) 35.4 3) 34.8 4) 36.3 1) 3.5 2) 3.3 3) 3.4 4) 3.4 F* 1) 3.4 2) 2.9 3) 3.5 4) 3.2 F** F x NDF x T** 1) 55 2) 53 3) 42 4) 29 Niagono et al., 1991 1) Control 2) Control + 1 kg yellow grease (Also included high and low degradability protein supplements) 1) 5.0 a 2) 16.3a 1-112 Ad libitum wheat silage and maximum of 16.4 kg/d of concentrate 1) 20.2 2) 19.0 1) 38.9 2) 39.8 1) 30.4 2) 29.4 1) 3.3 2) 3.4 1) 2.5 2) 2.4 1) -2.6 2) -24.4 F** Bateman et al., 1996 1) 33% NDF, 0% tallow 2) 33% NDF, 2% tallow 3) 40% NDF, 0% tallow 4) 40% NDF, 2% tallow 1) 3.5 2) 5.4 3) 3.7 4) 5.8 120-240, winter 8% alfalfa hay, 22% corn silage, 14% alfalfa silage, 0 or 13% earlage, 56 or 43% concentrate 1) 25.2 2) 22.6 3) 23.3 4) 24.3 F x NDF* 1) 31.1 2) 31.2 3) 28.6 4) 30.3 1) 30.3 2) 30.2 3) 28.1 4) 28.9 1) 3.30 2) 3.17 3) 3.22 4) 3.09 1) 3.94 2) 3.90 3) 3.96 4) 3.73 ------Bateman et al., 1996 1) 33% NDF, 0% tallow 2) 33% NDF, 2% tallow 3) 40% NDF, 0% tallow 4) 40% NDF, 2% tallow 1) 3.5 2) 5.4 3) 3.7 4) 5.8 120-240, summer 8% alfalfa hay, 22% corn silage, 14% alfalfa silage, 0 or 13% earlage, 56 or 43% concentrate 1) 21.7 2) 21.9 3) 22.7 4) 22.0 1) 29.4 2) 30.1 3) 30.7 4) 29.9 1) 26.9 2) 27.5 3) 29.8 4) 27.3 1) 3.14 2) 3.03 3) 3.25 4) 3.07 1) 3.52 2) 3.49 3) 3.86 4) 3.56 ------Son et al., 1996 1) 0% tallow + 0% escape protein supplement (EP) 2) 0% tallow + 5% EP 3) 3% tallow + 0% EP 4) 3% tallow +5% EP ------14-84 33% alfalfa haylage, 17% corn silage, 50% concentrate 1) 25.1 2) 24.2 3) 24.1 4) 22.8 F** F x EP** 1) 32.5 2) 33.0 3) 33.0 4) 33.1 1) 31.4 2) 30.1 3) 31.1 4) 31.7 F x EP** 1) 2.97 2) 2.80 3) 2.87 4) 2.75 1) 3.78 2) 3.41 3) 3.61 4) 3.71 1) -30.2 2) -20.3 3) -17.3 4) -37.1 F x EP** 3.5% FCM a concentrate mix only. F = effect of fat. T = effect of time. *P < .10. ** P < .05.

PAGE 51

38Table 2.3. Continued. Reference Treatments/ Fat source Dietary EE, % DIM Diet, % of DM DMI, kg/d Milk, kg/d 4.0% FCM, kg/d Milk protein, % Milk fat, % BW change, kg or kg/d Maiga et al., 1995 1) Control 2) 2% tallow 3) 8.3% tallow-molasses blend 4) 2% tallow + 5.4% whey 1) 2.9 2) 4.8 3) 4.7 4) 4.6 28-112 25% corn silage, 25% alfalfa hay, 50% concentrate 1) 23.1 2) 24.3 3) 24.5 4) 24.5 1) 31.9 2) 33.7 3) 33.7 4) 34.0 F** 1) 31.4 2) 33.9 3) 33.0 4) 33.4 1) 3.00 2) 2.98 3) 2.91 4) 2.86 2 vs. 3 & 4* 1) 3.48 2) 3.65 3) 3.52 4) 3.40 2 vs. 3 & 4* 1) 0.75 2) 0.42 3) 0.60 4) 0.61 (kg/d) F* Pantoja et al., 1996 1) Control 2) 5% tallow 3) 5% tallow + partially hydrogenated tallow (PHT) (2:1 weight/weight) 4) 5% tallow + PHT (1:2 weight/weight) 5) 5% PHT 1) 2.90 2) 6.13 3) 6.80 4) 6.58 5) 6.54 (FA) 28-133 25% corn silage, 25% alfalfa silage, 50% concentrate 1) 22.3 2) 22.1 3) 21.2 4) 22.5 5) 23.9 2 vs. 5* 1) 35.6 2) 40.6 3) 36.9 4) 39.3 5) 38.0 F** 1) 33.6 2) 35.6 3) 33.6 4) 36.3 5) 36.3 1) 3.05 2) 2.86 3) 3.03 4) 2.98 5) 3.02 2 vs. 5** 1) 3.63 2) 3.17 3) 3.48 4) 3.56 5) 3.77 2 vs. 5** 1) -2.16 2) -0.53 3) 1.09 4) 0.13 5) 0.04 (kg/wk) F** Salfer et al., 1995 1) Control 2) -14 0 DIM 1% partially hydrogenated tallow (PHT), 1-151 DIM 2% PHT 3) 1-151 DIM 2% PHT 4) 35-151 DIM 2% PHT Prepartum 1) 3.01 2) 3.92 Postpartum: 1) 3.17 2) 4.99 1-151 Prepartum: 37% corn silage, 10% alfalfa silage, 14% grass hay, 39% concentrate Postpartum: 24% corn silage, 21% alfalfa silage, 55% concentrate 1) 19.9 2) 19.9 3) 20.6 4) 20.6 1) 30.7 2) 31.6 3) 31.9 4) 32.7 1) 32.0 2) 32.7 3) 33.2 4) 32.8 1) 3.04 2) 2.99 3) 2.97 4) 2.92 1) 3.81 2) 3.76 3) 3.83 4) 3.55 N.S. Salfer et al., 1995 1) Control 2) -14 0 DIM 1% partially hydrogenated tallow (PHT), 1-151 DIM 2% PHT 3) 1-151 DIM 2% PHT Prepartum 1) 3.01 2) 3.92 Postpartum: 1) 3.17 2) 4.99 1-35 (3 treatments initiated) Prepartum: 37% corn silage, 10% alfalfa silage, 14% grass hay, 39% concentrate Postpartum: 24% corn silage, 21% alfalfa silage, 55% concentrate 1) 15.3 2) 15.5 3) 16.3 1) 30.2 2) 29.9 3) 30.6 1) 34.5 2) 33.4 3) 36.9 2 vs. 3* 1) 3.06 2) 3.08 3) 3.13 1) 4.45 2) 4.29 3) 4.81 2 vs. 3** N.S. 3.5% FCM. F = effect of fat. N.S. = not significant. P < .10. ** P < .05.

PAGE 52

39Table 2.3. Continued. Reference Treatments/ Fat source Dietary EE % DIM Diet, % of DM DMI, kg/d Milk, kg/d 4.0% FCM, kg/d Milk protein, % Milk fat, % BW change, kg or kg/d Salado et al., 2004 1) Control 2) 0.7 kg/d partially hydrogenated vegetable oil 1) 6.7 a 2) 22.5 a -15-75 Pasture and 5 or 4 kg of concentrate offered for treatments 1 and 2, respectively 1) 20.3 2) 19.7 1) 23.7 2) 25.0 F** 1) 22.5 2) 24.5 F** 1) 3.12 2) 3.14 1) 3.64 2) 3.86 F** 1) -0.40 2) -0.04 (kg/d) Markus et al., 1996 1) Control 2) 7.1% whole sunflower seeds 3) 2.7% tallow 1) 1.8 2) 4.2 3) 4.1 16-112 12% corn silage, 14% alfalfa silage, 9.5% alfalfa hay, 64.5% concentrate 1) 22.2 2) 21.1 3) 21.6 1) 34.4 2) 34.6 3) 35.5 1) 30.0 2) 29.9 3) 31.6 1) 3.1 2) 3.0 3) 3.0 1) 3.2 2) 3.1 3) 3.3 ------Weiss and Wyatt, 2003 1) Control 2) 12.3% whole roasted soybeans 3) 2.35% tallow (Also included 3 levels of vitamin E) ------160-188 38% corn silage, 8% alfalfa hay, 7% alfalfa silage, 47% concentrate 1) 22.3 2) 24.0 3) 22.0 2 vs. 3** 1) 35.1 2) 36.8 3) 37.5 F* ------1) 2.97 2) 2.92 3) 2.86 1) 3.76 2) 3.83 3) 3.08 F** 2 vs. 3** 1) 0.87 2) 1.29 3) 0.71 (kg/d) 2 vs. 3** Wu et al., 1994 1) Control 2) 12% WCS 3) 12% WCS + 2.2% safflower oil 4) 12% WCS +2.2% prilled tallow 5) 12% WCS +4.4% prilled tallow 1) 3.3 2) 5.2 3) 7.4 4) 7.4 5) 9.6 50-125 43% alfalfa hay, 57% concentrate 1) 28.2 2) 27.2 3) 28.8 4) 26.8 5) 24.1 1) 32.5 2) 32.6 3) 35.0 4) 34.3 5) 33.0 1 & 2 vs. 3 & 4** 1) 32.4 2) 32.3 3) 33.4 4) 34.6 5) 33.0 1 & 2 vs. 3 & 4** 1) 3.20 2) 3.03 3) 3.03 4) 3.08 5) 3.07 1 vs. 2 ** 1) 3.49 2) 3.48 3) 3.26 4) 3.58 5) 3.51 1) 0.70 2) 0.82 3) 0.57 4) 0.64 5) 0.46 (kg/d) Pires et al., 1996 1) Control 2) 18% ground roasted soybean 3) 18% whole soybean 4) 2.7% blood meal 5) 2.7% blood meal + 3% tallow 1) 3.2 2) 6.2 3) 6.2 4) 3.2 5) 6.2 21-126 30% corn silage, 20% alfalfa silage, 50% concentrate 1) 23.6a 2) 22.7ab 3) 21.3bc 4) 21.3bc 5) 20.4c 1) 39.6 2) 40.7 3) 36.4 4) 36.1 5) 39.3 1) 35.4 2) 35.0 3) 33.3 4) 33.9 5) 35.0 1) 3.03a 2) 2.83b 3) 2.88bc 4) 3.08a 5) 2.98ac 1) 3.33 2) 3.09 3) 3.50 4) 3.63 5) 3.29 ------a Concentrate mix only. WCS = whole cottonseeds. F = effect of fat. P < .10. ** P < .05.

PAGE 53

40Table 2.4. Effects of feeding ruminally inert fat (Ca salts of fatty acids (CSFA) or pril led fat (PF)) to the same cows through out the study on concentration of plasma hormones and metabolites. Reference Treatments/ Fat source Dietary EE, % DIM Diet, % of DM NEFA, eq/L BHBA, mg/dl Insulin, ng/ml IGF-1, ng/ml Other Erickson et al., 1992 1) Control 2) 12 g/d niacin (NA) 3) 3% CSFA 4) NA + CSFA ------15-98 35% alfalfa haylage, 10% corn silage, 55% concentrate 1) 265 2) 238 3) 303 4) 352 F** 1) 6.50 2) 5.18 3) 7.82 4) 6.48 F** ------------------Moallem et al., 2000 1) Control 2) 0.55 kg/d CSFA 3) Control + bST ------0-150 9.5% wheat silage, 15% corn silage, 3.5% legume hay, 3.5% oat hay, 2.3% wheat straw, 66.2% concentrate ------------------------Progesterone based cyclicity, DIM 1) 27 2) 29 3) 26 GarciaBojalil et al., 1998 1) 11.1% RDP 2) 11.1% RDP + 2.2% CSFA 3) RDP 15.7% 4) 15.7% RDP + 2.2% CSFA 1) 4.77 2) 6.65 3) 4.62 4) 6.20 0-120 34% corn silage, 13% alfalfa hay, 53% concentrate a RDP x F** a ------1) 0.69 2) 0.60 3) 0.55 4) 0.52 F* ------------Spicer et al., 1993 1) Control 2) 1.8% CSFA ------28-84 20% sorghum silage, 19% alfalfa hay, 61% concentrate a ------------------1) 36.1 2) 82.7 Progesterone 1) 6.03 2) 4.47 F** Selberg et al., 2004 1) Control 2) 225 g/d CaS conjugated linoleic acid (CLA) 3) 225 g/d CaS transC18:1 Control prepartum: 4.3 Control postpartum: 5.2 -28-49 Prepartum: 13% bermuda grass hay, 39% corn silage, 52% concentrate Postpartum: 10% alfalfa hay, 29% corn silage, 61% concentrate 1 vs. 2 by week** (CaS CLA peaked higher at wk 1 postpartum) 1 vs. 2 by week** (CaS CLA peaked higher at wk 1 postpartum) 1 vs. 3 at week 6** (CaS transC18:1 cows were greater) ------Mean hepatic lipid and TAG concentrations did not differ among diets a Concentrate mix included 6-15% whole cottonseed. b NEFA concentrations from 0-49 DIM were elevated for cows fed 11.1% RDP + CSFA in comparison to 11.1% DIP, while15.7% RDP + CSF A decreased concentrations in comparison to the 15.7% RDP group. From 0-14 DIM cows fed 15.7% RDP diet had elevated NEFA concentrations in comparision to 15.7% RDP + CSFA. F = effect of fat. P < .10. ** P < .05.

PAGE 54

41Table 2.4. Continued. Reference Treatments/ Fat source Dietary EE, % DIM Diet, % of DM NEFA, eq/L BHBA, mg/dl Insulin, ng/ml IGF-1, ng/ml Other Beam and Butler, 1998 1) Control 2) 2.59% PF 1) 4.8 2) 7.0 0-100 26% corn silage, 18% alfalfa haylage, 56% concentrate a 1) 554 2) 656 ------1) 0.37 2) 0.33 1) 47.7 2) 37.6 F* (1-3 wk PP) ------Jerred et al., 1990 c 1) Low silage (LS) 2) LS + 5% PF 3) Medium silage (MS) 4) MS + 5% PF 5) High silage (HS) 6) HS + 5% PF 1) 3.1 2) 6.5 3) 3.4 4) 7.2 5) 3.8 6) 7.2 5-105 Trt 1 & 2: 45% alfalfa silage, 55% concentrate Trt 3 & 4: 64% alfalfa silage, 36% concentrate Trt 5 & 6; 84% alfalfa silage, 16% concentrate ------1) 13.3 2) 19.1 F** ------------------Skaar et al., 1989 1) Control 2) 12 g/d niacin (NA) 3) 5% PF 4) 12 g/d NA and 5% PF 1) 3.4 b 2) 3.4 b 3) 11.8 b 4) 11.8 b -17-105 25% corn silage, 25% alfalfa silage, 50% concentrate 1) 298 2) 343 3) 320 4) 371 1) 10.8 2) 11.2 3) 10.2 4) 9.7 ------------Increase in total hepatic lipids by fat feeding at 0 DIM and at 5 wk postpartum (P < 0.15); hepatic TAG = not significant Pickett et al., 2003 1) Control 2) 500 ml/d propylene glycol (PG) drench 3) 0.45 kg/d CSFA drench 4) 500 ml/d PG + 0.45 kg/d CSFA drench 4.8 0-21 31% corn silage, 16% alfalfa hay, 9% alfalfa hay, 44% concentrate a Drenches were administered from 03 DIM 1) 643 2) 503 3) 602 4) 558 1) 10.8 2) 8.1 3) 9.4 4) 9.4 1) 0.66 2) 0.72 3) 0.63 4) 0.61 ------Hepatic TAG % at 7 DIM 1) 10.8 2) 6.1 3) 10.0 4) 10.0 a Concentrate mix included 6-15% whole cottonseed b concentrate mix only. c Note: there are no fat x forage interactions ; results are presented as 1) control (t reatments 1, 3 and 5) and 2) fat (treatme nts 2, 4, and 6). F = effect of fat. P < .10. ** P < .05.

PAGE 55

42Table 2.4. Continued. Reference Treatments/ Fat source Dietary EE, % DIM Diet, % of DM NEFA, eq/L BHBA, mg/dl Insulin, ng/ml IGF-1, ng/ml Other Schroeder et al., 2003 1) TMR fed control 2) Pasture + 6.7 kg/d corn based concentrate 3) Pasture + 6.7 kg/d concentrate with 0.8 kg CSFA 1) 4.5 2) 6.1 3) 8.1 117-152 Control: 59% corn silage, 41% concentrate Trt 2 and 3: ~84% pasture, 16% concentrate 1) 349 2) 289 3) 311 ------------------------Moallem et al., 1999 1) Control 2) 0.55 kg/d CSFA 3) Control + bST ------0-150 8% wheat silage, 20% corn silage, 3% pea hay, 3% oat hay, 66% concentrate a 1) 159 2) 152 3) 125 (measured only at 50 DIM) ------------------Progesterone, ng/ml 1) 33.0 2) 55.4 3) 30.0 F** Moallem et al., 1997 1) Control 2) 0.5 kg/d CSFA 3) Control + bST 4) 0.5 kg/d CSFA + bST ------0-150 10.4% wheat silage, 24% corn silage, 2.3% pea hay, 2.1% oat hay, 61.2% concentrate F** (fat feeding increased NEFA concentrations) N. S. ------------DIM to first ovulation 1) 24.5 2) 26.9 3) 28.4 4) 27.5 Sklan et al., 1994 1) Control 2) 2.5% CSFA 1) 2.8 2) 4.9 0-120 14.3% wheat silage, 15.3% corn silage, 6.4% vetch hay, 64% concentrate a 1) 153 2) 172 F x P* ------------------1st conception rate was lower for primiparous cows fed CSFA F x P** Cervantes et al., 1996 1) Control 2) 0.4 kg/d CSFA 3) 12 g/d nicotinamide (NM) 4) 0.4 kg/d CSFA + 12 g/d NM 1) 3.1 2) 5.1 3) 3.0 4) 5.0 112-150 Varied dependent on stage of lactation. Forages (35-60% of diet) utilized were alfalfa hay, alfalfa haylage, and corn silage. 1) 120 2) 157 3) 126 4) 151 F* 1) 3.9 2) 3.9 3) 3.8 4) 3.3 ------------------Sklan et al., 1991 1) Control 2) 2.6% CSFA ------0-120 13.7% corn silage, 11.3% vetch hay, 75% concentrate a Fat fed had greater concentrations until 40 DIM** ------------------------a Concentrate mix included 6-15% whole cotton seed. F = effect of fat. P = effect of parity. N.S. = not significant. P < .10. ** P < .05.

PAGE 56

43Table 2.5. Effects of feeding oilseeds alone or in combination with other fat sources to the same cows throughout the study on concentration of plasma hormones and metabolites. Reference Treatments/ Fat source Dietary EE, % DIM Diet, % of DM NEFA, eq/L BHBA, mg/dl Insulin, ng/ml IGF-1, ng/ml Other Drackley et al., 1998 1) Control 2) Control + 12 g/d niacin 3) 10% whole raw soybeans and 2.5% tallow 4) Treatment 3 + 12 g/d niacin 1) 2.75 2) 2.75 3) 6.04 4) 6.04 28-301 32.5% alfalfa haylage, 17.5% corn silage, 50% concentrate a 1) 98 2) 117 3) 134 4) 122 F** F x niacin** F x niacin x T** 1) 4.9 2) 4.8 3) 4.6 4) 5.3 F x T* F x niacin x T** ------------------Pires, et al., 1996 1) Control 2) 18% ground soybean 3) 18% whole soybean 4) 2.7% blood meal 5) 2.7% blood meal + 3 % tallow 1) 3.2 2) 6.2 3) 6.2 4) 3.2 5) 6.2 21-126 30% corn silage, 20% alfalfa silage, 50% concentrate 1) 197 2) 248 3) 200 4) 194 5) 233 ------------------------a After 175 DIM, diets were adjusted for decreased nutrient requireme nts. Forage content increased to 60% of DM. For treatments 3 and 4 whole raw soybeans were removed and tallow was decreased to 2.25% of the diet. F = effect of fat. T = effect of time. P < .10. ** P < .05.

PAGE 57

44Table 2.6 Effects of feeding rendered fats to the same cows throughout the study on concentra tion of plasma hormones and metabolites. Reference Treatments/ Fat source Dietary EE, % DIM Diet, % of DM NEFA, eq/L BHBA, mg/dl Insulin, ng/ml IGF-1, ng/ml Other Bateman et al., 1996 1) 33% NDF, 0% tallow 2) 33% NDF, 2% tallow 3) 40% NDF, 0% tallow 4) 40% NDF, 2% tallow 1) 3.5 2) 5.4 3) 3.7 4) 5.8 120-240, winter 8% alfalfa hay, 22% corn silage, 14% alfalfa silage, 0 or 13% earlage, 56 or 43% concentrate 1) 190 2) 220 3) 160 4) 210 F* ------------------------Bateman et al., 1996 1) 33% NDF, 0% tallow 2) 33% NDF, 2% tallow 3) 40% NDF, 0% tallow 4) 40% NDF, 2% tallow 1) 3.5 2) 5.4 3) 3.7 4) 5.8 120-240, summer 8% alfalfa hay, 22% corn silage, 14% alfalfa silage, 0 or 13% earlage, 56 or 43% concentrate 1) 450 2) 510 3) 460 4) 540 F* ------------------------Salado et al., 2004 1) Control 2) 0.7 kg/d partially hydrogenated vegetable oil 1) 6.7 b 2) 22.5 b -15-75 Pasture and 5 or 4 kg of concentrate offered for treatments 1 and 2, respectively 1) 659 2) 691 ------1) 0.52 2) 0.55 1) 140.1 2) 112.5 ------F = effect of fat. P < .10. ** P < .05.

PAGE 58

45 CHAPTER 3 EFFECTS OF THE TIMING OF INITIATI ON OF FEEDING CALCIUM SOAPS OF LONG CHAIN FATTY ACIDS ON PERIPA RTURIENT HOLSTEIN COWS DURING SUMMER Materials and Methods Cows and Diets Holstein cows, housed at the Dairy Res earch Unit near Hague, FL (29 44 N latitude, 82 26 W longitude) were bl ocked by predicted calving date, parity (primiparous or multiparous), body weight ( BW ), and milk production of the previous year for multiparous cows and assigned randomly to treatment at approximately 28 d prior to their due date. Parturition occu rred between April 24 and August 9, 2003 for all cows. A total of 58 cows were assigned to the experiment and calved having consumed their dietary treatment for at least 10 d prior to calving. Ho wever, due to lameness (n = 3), displaced abomasum (n = 3), death due to re spiratory infection (n = 1), severe mastitis (n = 1), severe vaginal tear (n = 1) ketosis, retained fetal membranes ( RFM ) and static rumen for 10 continuous d (n = 1), and an undiagnosed condition (n = 1), only 47 cows (n = 25 multiparous and n = 22 primiparous) were in cluded in the final data set. Sample collections, housing conditions, and animal car e met the requirements of the Animals Used for Teaching and Research Protocol approved by the Animal Care and Use Committee at the University of Florida, Gainesville. Animals were housed on pasture with shad e and cooling fans provided beginning at approximately 28 d prior to parturition. At approximately 7 d prior to parturition, cows were moved to a sand-bedded barn with coo ling fans and sprinklers to allow for close

PAGE 59

46 observation of signs of impendi ng parturition. After parturi tion cows were housed in an open-sided free stall barn equipped with sa nd bedding, cooling fans, sprinklers and selflocking stanchions where they were fed their assigned dietary treatment. Two diets were prepared and fed as a total mixed ration ( TMR ) (Table 3.1 and 3.2) in ad libitum amounts twice daily to allow for 5 to 10% refusal in the prepartum and postpartum periods, respectiv ely. Calcium salts of long chain fatty acids ( CSLCFA Megalac-R, Church & Dwight Co., Princeton, NJ ) were fed at 0 or 2.0% of dietary dry matter ( DM ). The fatty acid ( FA ) profile of CSLCFA provi ded by the manufacturer was 17.4% C16:0, 2.1% C18:0, 32.1% C18:1 cis 1.5% C18:1 trans 30.5% C18:2, 2.4% C18:3, and 12.2% other FA. Four experiment al treatments were the following: 0% CSLCFA (Control), CSLCFA fed starting at 28 d prepartum, CSLCFA fed starting at 1 d in milk ( DIM ), and CSLCFA fed starting at 28 DIM. Once initiated, all diets continued through 100 DIM. Those cows receiving CSLC FA beginning in the postpartum period were fed CSLCFA at 1% of dietary DM fo r 7 d in order to adapt cows to CSLCFA slowly. Sample Collection Representative samples were obtained of corn silage, bermudagrass hay, alfalfa hay, and concentrate mixes on a weekly basis. Corn silage was immediately dried at 55C for 48 h in a forced air oven in order to calculate concentration of DM and maintain the same formulated forage to concentrate ratio. The weekly samples were composited on a monthly basis, ground through a 1mm Wiley mill screen (A. H. Thomas, Philadelphia, PA) and analyzed for chemi cal composition using wet chemistry (Dairy One, Ithaca, NY). Cows were milked three times per day at 0200, 1000, and 1800 h. Milk production was recorded at each milki ng. Milk samples were collected weekly

PAGE 60

47 from two consecutive milkings and analyzed by Southeast Milk lab (Belleview, FL) for fat, true protein, and somatic cell count ( SCC ) using a Bently 2000 NIR analyzer. Body weight was measured weekly after the 0900 h milking. Body condition scores ( BCS ) were assigned by the same two individuals at -4, 0, 3, 6, 9, 12, and 14 wk postpartum (Edmonson et al., 1989). Blood wa s collected three times week ly from parturition until artificial insemination ( AI) (72 3 DIM) and again at 7 d post AI from the coccygeal or the jugular vessels immediately before th e 1000 h milking using 13 x 100 ml vacutainer tubes containing sodium hepari n (Becton Dickinson Vacutainer systems, Franklin Lakes, NJ). Samples were put imme diately on ice until centrifuged at 2619 g at 5C for 30 min (RC-3B refrigerated centrifuge, H 600A rotor, Sorvall Instruments, Wilmington, DE). Plasma was decanted and frozen at -20C. Liver samples were coll ected at 2, 14 1, and 28 1 DIM via liver biopsy, rinsed with ster ile saline and immediately frozen in liquid nitrogen (-192C) and st ored at -80C. Reproductive Management The previous pregnant uterine horn was determined as the longer horn with the greater diameter using a r eal time Ultrasound Aloka 500 s canner (Aloka Co., Ltd, Tokyo, Japan) equipped with a 5.0 MHz linear rectal transducer. The diameters of the uterine horn and cervix were measured at 21 3 and 28 3 DIM. The transducer was placed in a transverse position in relation to the horns, at approximately 4 cm past the bifurcation of the horns. When the transducer was posit ioned and the horns could be seen clearly, the image was fixed. Pressure with the tran sducer on the uterine horns was avoided in order to obtain a circular cross-section image of the horns. Machine calipers were activated such that a vertical line was extende d from serosa to serosa of the uterine horn cross-section. Cervical diameter was measur ed by placing the transducer in a transversal

PAGE 61

48 position in relation to the cervi x at its middle section and th e distance between two points was obtained as described above. A 7.5 MHz linear rectal transducer was used to monitor ovarian structures. Size, location and number of follicles were recorded at 21 and 28 DIM and categorized by size into Class 1 (2 to 5 mm), Class 2 (6 to 9 mm), and Class 3 ( 10 mm) follicles (Lucy et al., 1992). Th e location and size of corpus luteum ( CL ) (determined by measurement of le ngth and width) were recorded. At 21 3 and 28 3 DIM, uterine tonus was classified as none, moderate, and intense. Also, vaginal examinations were c onducted to characterize uterine condition at 21 3 and 28 3 DIM using a glass speculum in serted into the vagina until the cervical os could be seen. The cervix was classified as open or closed, th e cranial vagina was classified as pink or red, a nd vaginal discharge was classi fied according to amount (none, trace, slight, moderate, or copious) and quality (none, clear mu cous, cloudy mucous, mucopurulent, or purulent). Cows were enrolled in a Pre-synchronization/Ovsynch protocol beginning at 44 3 DIM by injecting gonadatrop in releasing hormone ( GnRH ) (100 g, Gonadorelin Diacetate Tetrahydrate, Cystorelin, Merial Ltd., Athens, GA) and ovaries were scanned every other day for 7 d. At 51 3 DIM, prostaglandin F2 ( PGF2 ) (25 mg, Lutalyse, Pharmacia Upjohn, Kalamazoo, MI) was injecte d. Ultrasound examination of the ovaries continued until the formation of a new CL. Eleven days later (62 3 DIM), GnRH was injected again (100 g), followed by PGF2 (25 mg) 7 d later (69 3 DIM), and GnRH (100 g) 48 h following PGF2 (71 3 DIM). Ovaries we re scanned by ultrasonagraphy at each injection day. All hormone injections were given intramuscularly (i.m.) after a blood collection from coccygeal vessels. Co ws were inseminated at 72 3 DIM (16 h

PAGE 62

49 following GnRH) using the same batch of semen by the same trained inseminator. Immediately after AI, an injection of bST (POSILAC Monsanto Co., St. Louis, MO) was given in the ischiorectal fossa and continued every 14 d thereafter. At 7 d post AI (79 3 DIM), ovaries were examined by ultrasound for the presence of a CL. Cows with no CL were reenrolled in Ovsynch. At 28 d post AI (100 3 DIM), pregnancy was determined by identifying the presence of embryonic fluid, appearance of embryo and an embryonic heart beat using u ltrasound. Pregnancy was confir med by rectal palpation at 45 and 72 d post AI by the farm veterinarian. Animal Health Body temperature was measured the fi rst 5 d postpartum using a rectal thermometer. Cows were diagnosed as havi ng RFM if membranes we re still attached 24 h after parturition. Treatment of RFM was peni cillin (50 cc, i.m.) for 5 d. Cows having a displaced abomasum had corrective surgery pe rformed by the farm veterinarian, followed by administration with dextrose (500 ml) a nd vitamin B complex (15 cc) intravenously plus calcium propionate (300 ml) and 30 g of Pr obios (Vets Plus Inc., Knapp, WI) orally. Ketosis was diagnosed using Ketostix (Bayer, Pittsburgh, PA). Treatment for ketosis was dextrose (500 ml) and vitamin B complex (15 cc) intravenously and 30 g of Probios (Vets Plus Inc., Knapp, WI) orally. Cows having a rectal temper ature > 39.4C and with no other apparent health conditi ons were diagnosed as having metritis and treated with 0.23 to 0.45 mg/kg BW of Nexcel (ceftiofur s odium sterile powder, Pfizer Inc., New York, NY). Sample Analysis Plasma concentrations of nonesterified fatty acids ( NEFA ) (NEFA-C kit; Wako Fine Chemical Industries USA, Inc., Dalla s, TX; as modified by (Johnson, 1993) and -

PAGE 63

50 hydroxy butyric acid ( BHBA ) (Pointe Scientific Inc., Linc oln Park, MI) were determined once weekly. A Technicon Autoanalyzer (T echnicon Instruments Corp., Chauncey, NY) was used to determine weekly con centrations of bl ood urea nitrogen ( BUN ) (a modification of (Coulombe and Favreau, 1963) as described in Bran + Luebbe Industrial Method #339-01) and plasma glucose (a modi fication of (Gochman and Schmitz, 1972) as described in Bran + Luebbe Industrial Method #339-19). A double antibody radioimmunoassay ( RIA ) was used to determine plasma concentrations of insulin (B adinga et al., 1991; Malven et al., 1987) and IGF-1 (Badinga et al., 1991) on every plasma sample collected. The sensitivity of the insulin assay was 0.3 ng/ml, and intraand interassa y coefficients of variation ( CV ) were 9.1 and 14.9%, respectively. The sensitivity of the IGF-1 assay was 50 pg/ml, and in traand interassay CV were 11.4 and 12.1%, respectively. Weekly leptin concentrations were determined by RIA at the University of Missouri (Delava ud et al., 2000). Intraand interassay CV were less than 10%. Concentr ations of plasma progesterone were determined on every plasma sample collected using Coat-A-Count Kit (DPC Diagnostic Products Inc., Los Angeles, CA) solid phase 125I RIA. The sensitivity of the assay was 0.1 ng/ml and the intraand interassay CV were 0.8 and 5.2% respectively. A polye thylene glycol RIA procedure described by Meyer et al. (1995) was used to anal yze the concentration of 15keto-13,14-dihydro-prostaglandin F2 ( PGFM ) on daily plasma samples collected during the first 10 DIM. The sensitivity of the assay was 31.25 pg/ml and the intraand interassay CV were 8.2 and 20.6%, respectively. The three plasma samples collected week ly were composited into one sample during each of the first 4 wk postpartum a nd analyzed for concentrations of alkaline

PAGE 64

51 phosphatase ( ALK ) (Diagnostic Chemicals Ltd, Oxford, CN), alanine aminotransferase ( ALT ) (Abbott Diagnostics, Abbott Park, IL ), aspartate aminotransferase ( AST ) (Abbott Diagnostics, Abbott Park, IL), gamma glutamyl transferase ( GGT ) (Thermo DMA, Arlington, TX), albumin (Diagnostic Chemical s Ltd, Oxford, CN), and total biliruben (Wako Chemicals USA, Inc., Richmond, VA) using a Hitachi 911 chemistry analyzer (Roche Diagnostics, Indianapolis, IN). Acute phase protein concentrations were determined on two samples per week for 4 wk postpartum. Plasma fibrinogen was dete rmined from a standard curve generated using a human reference (Sigma Diagnostics). Plasma haptoglobin concentrations were determined by measuring haptoglobin/hem oglobin complexing (Makimura and Suzuki, 1982). Ceruloplasmin oxidase activity was m easured using colorimetric procedures described by Demetr iou et al. (1974). Total liver cellular RNA was isolated fr om samples collected at 2, 14, and 28 DIM from eight cows in the control, fat pre, and fat 1 DIM groups (n = 72) using TRIzol reagent (Life Technologies, Grand Isla nd, NY) according to the manufacturers directions. Ten g of R NA was fractionated in a 1.0% agrose-formaldehyde gel and transferred overnight to Bi oTrans nylon membranes (ICN, Irvine, CA) by capillary action. The RNA was cross linked to the memb rane by exposure to u ltraviolet light for 90 sec and the membrane was heated at 80C for 1 h. The membranes were prehybridized for 30 min with buffer (ULTRAh yb buffer, Ambion Inc., Austin, TX) to block non-specific binding sites. Me mbranes were then hybridized to 32P-labeled IGF-I, IGF-II, and IGFBP-2 cDNA probes, respectiv ely. After hybridization, filters were washed for 15 min in 50 ml of 2X SSC, 0.1% SDS at 50C, followed by two 15-min

PAGE 65

52 washes in 50 ml of 0.1X SSC, 0.1% SDS at 50C. Filters were blotted dry and exposed to x-ray film (Super RX, Fugi Film, Japan) fo r 3 to 96 h at -80C. Hybridization signals were quantified by densitom etric analysis (Eastman Kodak, Rochester, NY). Triacylglycerol ( TAG ) concentrations in liver samp les (75 to 150 mg, wet weight) were determined colorimetri cally (Foster and Dunn, 1973) by first extracting the total lipid as described by Drackley et al. (1992). Dry matter c ontent of liver (~100 mg) was determined by drying in a forced air oven for 48 h at 55C. Statistical Analysis Data were analyzed as a completely randomized design using the PROC MIXED procedure for repeated measurement of SAS (S AS software statistics, 2001) according to the following mathematical model: Yijkl = + Di + Pj + DPij +Ck (i j) +Wl + DWil + PWjl + DPWiikl + Eijkl where Yijkl is the observation, is the overall mean, Di is the fixed effect of diet (i = 1, 2, 3, and 4), Pj is the fixed effect of parity (j = 1 and 2), DPij is the interaction of diet and parity, Ck (i j) is random effect of cow within diet and parity (k = 1, 2, n), Wl is the fixed effect of week (l = 0, 1, 2, ), DWil is the interaction of diet and week, PWjl is the interaction of pa rity and week, DPWijl is the three way interac tion of diet, parity and week, and Eijkl is the residual error. Results are reported as least square m eans. Significance was determined at P 0.05 and tendencies included P > 0.05 and 0.10. Orthogonal contrasts used included 1) no CSLCFA versus CSLCFA (control vs. (CSL CFA pre, CSLCFA 1 DIM, plus CSLCFA 28 DIM)), 2) CSLCFA feedi ng initiated prepartum vs. CSLCFA feeding initiated postpartum (CSLCFA pre vs. (CSLCFA 1 DIM plus CSLCFA 28 DIM)), and 3) CSLCFA initiated at 1 DIM versus CSLCFA initiated at 28 DIM. These three contrasts

PAGE 66

53 by parity interactions were tested also. In addition, for dependent variables measured only before 28 DIM, the cows assigned to the CSLCFA 28 DIM treatment were combined with control cows. For this data set (when only three treatments had been initiated), a separate set of orthogonal contrasts was used. Contrasts were 1) CSLCFA feeding initiated in the prepartum vs. no CSLCFA feeding in the prepartum period (CSLCFA pre vs. (Control plus CSLCFA 1 DIM plus CSLCFA 28 DIM)), and 2) no CSLCFA (Control plus CSLCFA at 28 DIM) vs CSLCFA initiated at 1 DIM. These two contrasts by parity interac tions were tested also. Data for NEFA, BHBA, BUN, glucose, leptin, alkaline pho sphatase, alanine aminotransferase, aspartate aminotransferase, gamma glutamyl transferase, albumin, total biliruben, hepatic mRNA, hepatic TAG, m ilk production, BW, and BCS were evaluated using repeated measures of the Mixed procedur e of the SAS software program. Data were tested to determine the structure of best fit, namely AR (1), ARH (1), CS, or CSH, as indicated by a lower Schwartz Baesian inform ation criterion value (L ittell et al., 1996). Orthogonal contrasts (cited above ) were used to determine if the mean values were different. If main effects or contrasts were significant, the slice command was used to determine at which time points the treatments di ffered. If slice was significant, then the pdiff command was used to determine which trea tments were different at that time point. Data that did not have a common day in milk (insulin, IGF-I, P4, PGFM, fibrinogen, haptoglobin, and cer uloplasmin) were modeled us ing the CS structure as a polynomial function of time using regression anal ysis and coefficients were obtained to plot the curves after the leve l of order (linear, quadratic, c ubic, quartic, or quintic) that

PAGE 67

54 best fit the data was determined. Hete rogeneity of regression was performed to determine if the curves differed for each of the orthogonal contrasts cited above. Data for average milk protein concentr ation, average milk fat concentration, average milk protein yield, average milk fat yield, 3.5% fat corrected milk ( FCM ) yield, vaginal exams, and 21 DIM and 28 DIM ovarian maps were analyzed using the Proc GLM procedure of SAS. Odds ratios and their confidence intervals were obtained using the Chi square and PROC LOGISTIC procedures for testing of first service conc eption rates, estrus cycling rates (the first day postpartum of ovulat ion was determined by plasma progesterone concentrations > 1 ng/ml for two consecutive sampling days) and incidence of disease. Results and Discussion Dry Matter Intake and Diets Concentrations of nutrients in experime ntal diets were within or exceeded the acceptable targets for close-up nonlactating cows and lactating cows at 90 DIM (NRC, 2001) (Tables 3.1 and 3.2). The ether extract values of the diets containing CSLCFA increased 1.3 and 1.6 percentage units for th e close-up and lactati ng diets respectively. During the prepartum period, cows consum ed an average of 10.0 and 10.2 kg of DM/d when offered the 0 or 2% CSLCFA diet s, respectively. Cows were fed diets an average of 24.8 7.4 d prior to parturition. During the postpartum period, cows consumed an average of 17.5 and 16.4 kg of DM/d when offered the 0 or 2% CSLCFA diets, respectively. In a revi ew of several studies, Allen ( 2000) reported a linear decline in DMI of 2.5% for every 1% inclusion of CSLCFA in the diet DM. Although not statistically evaluated, the diffe rence of 6.3% is close to th e 5% depression of intake predicted by Allens (2000) equations.

PAGE 68

55 Production and Body Weight As expected, multiparous cows were heavier (644 vs. 530 kg; P = 0.001) and produced more milk (43.4 vs. 41.0 kg/d; P = 0.041) than primiparous cows (Table 3.3). In addition, multiparous cows tended to have a greater SCC than did primiparous cows (586 vs. 354 x 1000/ml; P = 0.067; Table 3.3), however no treatment differences were detected. It is well documented that milk producti on often increases when supplemental fats are fed. This may be due to increased ener gy intake. In the current study, cows fed CSLCFA in the prepartum period tended to produce more milk (42.2 vs. 37.1 kg/d; P = 0.059) and 3.5% FCM (41.3 vs. 36.5 kg/d; P = 0.087) the first 100 DIM than cows fed CSLCFA beginning in the pos tpartum period (Table 3.3). Feeding CSLCFA beginning prepartum probably increased milk producti on due to increased energy intake and because the cows did not have the additio nal stress of adjusting to a new dietary ingredient early in lactation. Frequently there is a three to four wk delay before milk production increases when supplemental fats are added during the postpartum period (Garcia-Bojalil et al., 1998; Schingoethe and Ca sper, 1991). In contrast to the results of this study, Salfer and coworkers (1995) did not detect any differenc e in milk yield or 3.5% FCM yield of cows fed diets of 0 or 2% partially hydrogenated tallow ( PHT ) beginning at 14 d prepartum, at 1 DIM, or at 35 DIM through 151 DIM. However, when only the first 35 DIM were considered, yiel d of 3.5% FCM by cows fed PHT beginning at 1 DIM tended to be greater than that of cows fed PHT beginning 14 d prepartum (33.4 vs. 36.9 kg/d). Milk protein concentration wa s reported to not change (A twal et al., 1990; Markus et al., 1996; Moallem et al., 2000) or to d ecline when supplemental fats were fed

PAGE 69

56 (AbuGhazaleh et al., 2004; Cervantes et al ., 1996; Drackley et al., 1998). Protein concentration in the milk of primiparous cows was 2.63% for both no CSLCFA and CSLCFA fed groups, however multiparous cows fed CSLCFA had increased milk protein concentration in comparison to cows not fed CSLCFA (2.70 vs. 2.47%) (no CSLCFA vs. CSLCFA by parity interaction, P = 0.091). Th is increase could be due to more ruminal microbes delivering protein for protein synt hesis by the mammary gland. When fat is fed, bacteria numbers may increase in c oncurrence with a decrease in protozoal populations (Sutton et al., 1983). Likely due to increased milk production, milk protein yield tended to be greater by cows fed CSLC FA prepartum in comparison to cows fed CSLCFA beginning postpartum (1.15 vs. 1.02 kg/d; P = 0.08). In this study, no treatment or parity differences were detected for average milk fat concentration (3.44%) or yield (1.32 kg/d) through 100 DIM. In contrast, some studies reported an increase in milk fat concentrati on when CSLCFA were included in the diet (Cervantes et al., 1996; Moallem et al., 2000; Sklan et al., 1994; Sk lan et al., 1991). The response of milk fat concentration to dietary fat is depe ndent upon many factors including the fat concentration and composition in the basal diet and in the supplement as well as the forage source and amount. The ru minally inert fat fed in this study should not have interfered with ruminal digestion and the diet included sufficient fiber from both corn silage and alfalfa hay. About 50% of the fat found in milk is synthesized in the mammary gland from acetate and butyrate, whil e the other 50% comes directly from fat absorbed from the blood (Ackers, 2002). Increased uptake of long chain FA by the mammary gland for milk fat synthesis may i nhibit the synthesis of short and medium chain FA (Bauman and Griinari, 2003). Becau se we did not analyze the FA present in

PAGE 70

57 the milk, we can only speculate that there was a balance between increased dietary FA uptake and decreased synthesis of short a nd medium chain FA in the mammary gland such that total concentration of milk fat di d not differ from control cows. Palmquist and coworkers (1993) published an equation based on 49 published experiments predicting an increase in milk fat concentration of 0.18% due to feeding an additional 0.5 kg of fat daily. In the present study, milk fat concen tration was 0.11% units greater for cows fed CSLCFA versus controls however this was not significantly different. In agreement with several other studies in which fat supplementation was initiated in the periparturient period (G arcia-Bojalil et al., 1998; Ki m et al., 1993; Moallem et al., 2000), treatment did not affect mean postpartu m BW (587 kg). Multiparous cows not fed CSLCFA continued to gradually lose BW fo r 12 wk whereas those fed CSLCFA reached nadir by two to three wk postpartum. The pa ttern of BW for primiparous cows appeared similar for all treatment groups (treatme nt by parity by week interaction, P < 0.001, Figure 3.2). A treatment by week interacti on was detected for body condition score (P = 0.007, Figure 3.3). Cows fed CSLCFA beginni ng at 1 or 28 DIM reached their nadir sooner and began gaining body condition earlier postpartum than controls or cows fed CSLCFA beginning prepartum. Greater loss of body condition by thes e latter two groups of cows was most likely due to less energy in take and greater milk yield respectively. Plasma Metabolites Typically, plasma BHBA concentrations are not affected by fat supplementation, but can decrease slightly in fat-supplemented cows if cows on the basal diet have high BHBA concentrations (Grummer and Carro ll, 1991). In this study, mean plasma concentration of BHBA was lower for cows fed CSLCFA in comparison to controls (8.7 vs. 12.5 mg/dl, respectively; P = 0.007; Table 3.4). This difference was particularly

PAGE 71

58 apparent for control versus CSLCFA 1 DI M and 28 DIM at wk 3, 4, and 5 postpartum and at wk 5 for CSLCFA prepartum (P < 0.05; Figure 3.4). Control cows may have been mobilizing more of their adi pose tissue early in lactati on to support milk production due to the lower energy dense diet than cows fe d CSLCFA. In addition, an interaction of treatment and parity was detected (Table 3.4). Multiparous cows fed CSLCFA had lower concentration of plasma BHBA than control cows whereas th at of primiparous cows was greater but unaffected by di et (no CSLCFA vs. CSLCFA by parity interaction, P = 0.044). When fat is supplemented, plasma concentrations of NEFA routinely increase (Drackley, 1999), although ther e are several reports of num eric increases in NEFA concentrations that are not significant (Beam and Butler, 199 8; Salado et al., 2004). In this experiment, control cows did not diffe r from cows fed CSLCFA, probably due to control cows and CSLCFA prepartum follo wing a similar pattern while cows fed CSLCFA beginning at 1 DIM and 28 DIM were si milar but followed a different pattern. Mean plasma concentrations of NEFA we re greater for cows fed CSLCFA in the prepartum period in comparison to cows fe d CSLCFA beginning postpartum (456 vs. 294 meq/L; P = 0.002). This difference was particul arly evident at wk 3 and 4 postpartum (P < 0.05, Figure 3.5). A large portion of the plas ma NEFA in cows fed fat prepartum may have been from adipose mobilization to s upport greater milk production because plasma BHBA followed a similar pattern although not si gnificant. Grum et al. (1996) reported lower concentrations of plasma NEFA postp artum when cows were fed supplemental fat (Qual-Fat) beginning prepartum in compar ison to cows not fed supplemental fat prepartum.

PAGE 72

59 Concentrations of plasma glucose are not affected generally by fat supplementation (Grummer and Carroll, 1991). However in th is study, concentration of plasma glucose was increased in cows fed CSLCFA in comp arison to controls (65.6 vs. 60.4 mg/dl; P = 0.004) but this was observed mainly in multiparous cows. Similar to what was observed with BHBA, multiparous cows not fed CSLCFA tended to have lower concentrations of plasma glucose than those fed CSLCFA (55.2 vs. 65.5 mg/dl) but primiparous cows across diets did not differ in plasma gluc ose (no CSLCFA vs. CSLCFA by parity interaction, P = 0.054). Elevated blood ketone s are associated often with lowered blood glucose. In addition, concentration of plas ma glucose was lower for cows fed CSLCFA beginning prepartum versus postpartum ( 62.8 vs. 67.0 mg/dl; P = 0.025) and was greater for primiparous cows versus multiparous cows (66.8 vs. 61.7 mg/dl; P = 0.001). A tendency for an interaction of treatment and week was detected (P = 0.10) and treatment differences were evident at wk 3, 4, 5, and 6 postpartum (P < 0.05; Figure 3.6). Control cows had lower concentrations of plasma gl ucose than those fed CSLCFA at 1 DIM and 28 DIM at wk 3, 4, and 5 postpartum, than those fed CSLCFA prepartum at wk 5 postpartum, and than those fed CSLCFA at 1 DIM at wk 6 postpartum. Cows fed CSLCFA prepartum had lower plasma concentr ations of glucose than those fed CSLCFA at 28 DIM at wk 3 and lower than those fed CSLCFA at 1 DIM at wk 6 postpartum. Differences in plasma glucose concentrations may be due to differences in DMI and milk production between treatments and parities. In a review of dietary fat and adipose tissue metabolism, Chilliard (1993) noted that in 52 comparisons the difference in plasma glucose between control and fat-supp lemented groups was 0 g/L ( 0.03).

PAGE 73

60 Mean plasma concentrations of BUN tended to be greater for cows fed CSLCFA starting prepartum in comparison to cows fed CSLCFA starting postpartum (12.9 vs. 10.6 mg/dl; P = 0.053). This was especially true for multiparous cows (15.3 vs. 10.9 mg/dl) compared to primiparous cows (10.4 vs. 10.2 mg/dl) (CSLCFA prepartum vs. CSLCFA postpartum by parity interaction, P = 0.081). A tendency for a treatment by week interaction was detected (P = 0.06; Figure 3.7), however multiple means contrasts did not detect any differences among means. Indivi dual DMI were not measured however milk production was greater by cows fed CSLCFA beginning prepartum, so we can speculate that cows fed CSLCFA beginning prepartum were consuming more DM and therefore more dietary protein than cows fe d CSLCFA beginning postpartum. Plasma Hormones Leptin is a hormone synthesized by adipos e tissue that is stimulated by adiposity and inhibited by undernutrition. Concentrations of leptin we re decreased around the time of parturition in concurrence with negativ e energy balance and a reduction in adipose stores and may have been mediated by the re duction in plasma insulin (Block et al., 2003). Primiparous cows fed CSLCFA tended to have a lower concentration of plasma leptin in comparison to controls (2.15 vs. 3.06 ng/ml) whereas multiparous cows fed CSLCFA tended to have greater concentrations of leptin in compar ison to controls (2.39 vs. 1.65 ng/ml; Table 3.4; Figure 3.8) (no CSLC FA vs. CSLCFA by parity interaction, P = 0.078). Low concentrations of circulating plasma leptin were correlated highly with greater milk production (Liefers et al., 2003) This relationship appears to fit with animals not fed CSLCFA; that is the higher producing multiparous cows had lower plasma concentrations of leptin compared to the lower producing primiparous cows (1.65 vs. 3.06 ng/ml). However when CSLCFA we re included in the diet, this inverse

PAGE 74

61 relationship disappeared. In creasing the triglyceride con centration of the blood may possibly influence leptin synthesis and release. Insulin-like growth factors play a dive rse role physiologically and are very dependent upon the nutritional state of the an imal. Plasma IGF-I concentrations are correlated positively with body condition and DMI. During the periparturient period when cows are often in a negative energy bala nce, circulating concen trations of IGF-I are low (Vega et al., 1991). As the cow continues through lactation and a positive energy status is restored, circulating concentrat ions of IGF-I will increase. Low IGF-I concentrations were associated with an exte nded postpartum interval to estrus in beef cows and also with delayed puberty (Roberts et al., 1997; Rutter et al., 1989), indicating that IGF-I can be correlated positively w ith reproductive performance. Insulin-like growth factor I acts synergistically with lu teinizing hormone (LH) to promote follicular development (Lucy, 2001). Beam and Butler (1 998) reported lower mean concentrations of plasma IGF-I (37.6 vs. 47.7 ng/ml) from wk 1 to 3 postpartum in lactating dairy cows fed a diet of 2.6% prilled fat compared to no supplemental fat despite no differences in energy balance. However, other studies re ported no differences in concentration of plasma IGF-I when supplemental fat was fed (S alado et al., 2004; Spic er et al., 1993). In the present study, primiparous cows had a greater mean concentration of IGF-I in comparison to multiparous cows when consuming the control diet (77.1 vs. 54.3 ng/ml); however when CSLCFA were fe d, primiparous cows had a lower mean concentration (77.1 vs. 64.2 ng/ml) whereas concentration of IGF-I increased in multiparous cows (54.3 vs. 68.2 ng/ml) (interac tion of treatment and parity; P = 0.040) (Table 3.4). Concentrations of IGF-I were lowest im mediately postpartum for all

PAGE 75

62 treatments and rose throughout the lactation period. The qua dratic pattern of plasma IGF-I concentration over time was different fo r animals fed the control diet and animals fed CSLCFA (P < 0.01; Table A-1) and fo r animals fed CSLCFA beginning prepartum versus beginning postpartum (P < 0.01; Table A-1). Likewise, the pattern over time was different for cows fed the control diet and cows fed CSLCFA by parity (P < 0.05; Table A-4) and for cows fed CSLCFA beginning prepartum versus beginning postpartum by parity (P < 0.01; Table A-4). Primiparous cows fed no CSLCFA experienced a more rapid increase in concentrations of plasma IGF-I after partur ition than CSLCFA-fed cows before reaching a plateau at 49 DIM; howev er plasma IGF-1 of cows fed CSLCFA continued to rise over time (Figure 3.9A). At 77 DIM there was no difference in concentrations of plasma IGF-I between the two treatment groups. Multiparous cows fed CSLCFA had greater plasma IGF-I concentra tions at parturition than cows not fed CSLCFA and their concentrations rose steadily over time so that at 77 DIM, cows fed CSLCFA had a much greater concentrati on than control cows (Figure 3.9B). Concentrations of plasma IGF-I of primip arous cows fed CSLCFA beginning prepartum versus postpartum differed little over time (F igure 3.10A). However, multiparous cows fed CSLCFA beginning postpartum had greater concentrations of IGF-I at parturition and rose slightly but steadily over time whereas those fed CSLCFA beginning prepartum rose at a greater rate starting at 42 DIM to surp ass cows fed CSLCFA beginning postpartum at 77 DIM (Figure 3.10B). The differences in ci rculating IGF-I concen trations may have been due to differences in energy balance. Fat supplementation has had mixed results on circulating concentration of plasma insulin (Staples et al., 1998). Florida workers (Garcia-Bojalil et al., 1998) reported

PAGE 76

63 decreased insulin con centrations when periparturient cows were supplemented with CSLCFA at 2.2% of dietary DM, however othe rs reported no difference between fat-fed cows and controls (Beam and Butler, 1998; Salado et al., 2004). Insulin has also stimulated ovarian follicle cell growth. Wh en granulosa cells from small (1 to 5 mm) follicles were cultured, the addition of insulin increased cell numbers several fold and increased progesterone production in comparis on to the control (La nghout et al., 1991). In the present experiment, plasma insulin co ncentrations increased gradually as week postpartum increased (Figure 3.11), however no treatment or parity effects for mean plasma concentration of insulin were detected (0.65 ng/ml, Table 3.4). Over time, eight individual quadratic curves for each trea tment by parity combination fit the data significantly better than one pooled curv e (Figure 3.11). However the orthogonal contrasts for treatment, parity, and treatmen t by parity were not significant for plasma concentrations of insulin over time (Tab le A-4). Although ther e were no treatment differences, it is noteworthy that plasma in sulin and glucose concentrations followed a similar pattern. Dietary fats typically increase concentra tions of circulating cholesterol, the precursor of progesterone (Grummer and Ca rroll, 1991). Ruminants fed supplemental fat often have a slight increase in blood progesterone concentrat ion (Staples et al., 1998). Progesterone, secreted by the CL, prepares the uterus for implantation of the embryo and helps maintain pregnancy by providing a nourishi ng environment for the conceptus. At breeding, greater concentrations of plasma pr ogesterone has been associated with higher conception rates (Butler et al ., 1996). Work by Hawkins et al. (1995) suggested that the increase seen in circulating progesterone wh en cows were fed supplemental fat was from

PAGE 77

64 a reduced rate of clearance of progesterone rather than from an increased synthesis of progesterone. Son et al. (1996) reported greater blood cholestero l and peak plasma progesterone concentrations duri ng the second ovulatory cycle in cows fed tallow at 2 vs. 0% of dietary DM accompanied by a tendency of improved conception. Workers at the University of Florida (Garcia-Bojalil et al., 1998) reported that accumulated plasma progesterone from 0 to 50 DIM was greater, pregnancy rates improved, and energy status did not change when cows were fed diets of 2.2% calcium salts of palm oil compared to non fat-supplemented cows. The cubic pattern over time of plasma accumulated progesterone was different between parities when cows were fed the control versus the CSLCFA-supplemented diet (P < 0.05; Table A-4). Primiparous cows fe d CSLCFA had a slightly greater rise in plasma concentration of accumulated proge sterone beginning at 25 DIM compared to cows fed no CSLCFA whereas multiparous co ws fed no CSLCFA had a slightly greater rise in plasma concentration of accumulated progesterone beginning at 23 DIM than cows fed CSLCFA (Figure 3.12). Cows were inse minated at 72 3 DIM when there was very little difference in accumulated progesterone concentrations among treatment and parity groups. Patterns over time were detected to be different between parities when fed CSLCFA beginning at 1 DIM or at 28 DI M (P < 0.01; Table A-4). Upon closer examination of the data, two primiparous co ws fed CSLCFA beginning at 1 DIM did not ovulate until 40 and 60 DIM (determined by two consecutive plasma progesterone concentrations > 1.0 ng/ml) which could ha ve skewed the accumulated progesterone concentrations of the group. To determine if differences existed before cows were enrolled in a synchronization program, accumu lated plasma progesterone concentrations

PAGE 78

65 before the first hormone injection (46 DIM 3) were analyzed. The proportion of cows that ovulated before the first GnRH inj ection (44 3 DIM) we re 91% (10/11), 75% (9/12), 83% (10/12) and 100% (12/12) for control, CSLCFA prepartum, CSLCFA at 1 DIM, and CSLCFA at 28 DIM respectively and did not differ. The DIM at first ovulation (Table 3.4) did not differ among treatments e ither (mean of 27 DIM). The quadratic pattern over time differed for accumulated plas ma progesterone concentrations of cows fed the control diet and cows fed CSLCFA by parity when only 1 to 46 DIM were included (P < 0.01; Table A-4; Figure 3.13) ; however patterns over time were not different between cows fed CSLCFA beginni ng at 1 DIM and at 28 DIM (Table A-4). We concluded that the difference in accumu lated progesterone over time between cows fed CSLCFA beginning at 1 DIM and at 28 DIM was not different but was due to two animals that experienced delayed ovulation. Through a series of desaturase s and elongases, linoleic acid ( LA ) (C18:2) can form dihomo-linolenic acid, a direct precursor to the series 1 prostaglandins, or can be further desaturated to arachidonic acid (C20: 4), a direct precursor to the 2 series prostaglandins (Biochemistry of Lipi ds, Lipoproteins, and Membranes, 1996). Prostaglandin F2 synthesized by endometrial tissue is an important regulator of parturition and the estrou s cycle by causing regression of the CL. Immediately prepartum, PGF2 is important in regressing the CL of pregnancy and circulating PGF2 concentrations decline as the postpartum uter us declines in size. Concentrations of PGFM may be associated with immune f unctions such as cellular immunity and neutrophil function. If LA is supplemented in the diet prepartum, more arachidonic acid

PAGE 79

66 may be synthesized leading to higher concentr ations of the series 2 prostaglandins and possibly a greater immune competence. Using heterogeneity of regression, the cubi c patterns of plasma concentrations of PGFM over time were different for cows fed CSLCFA prepartum in comparison to cows not fed CSLCFA prepartum (P < 0.05, Table A-2). However parity status also influenced this contrast (parity by no CSLCFA prepar tum vs. CSLCFA prepar tum interaction, P < 0.05, Table A-5). Primiparous cows fed CSLC FA prepartum had similar initial plasma PGFM concentrations as primiparous cows not fed CSLCFA prepartum, however peak concentrations (at ~4 DIM) were greater fo r cows fed CSLCFA prepartum and declined at a slower rate (Figure 3.14A). Multipar ous cows not fed CSLCFA prepartum had high initial plasma PGFM concentrations, continue d to decline and stab ilized at 10 DIM. Multiparous cows fed CSLCFA prepartum had peak concentrations of plasma PGFM at 4 DIM, slowly declined and st abilized at 12 DIM (Figure 3.14B). The reason why there was an interaction of treatment and parity is unclear. When cows that experienced RFM and/or metritis were excluded from this analys is, the patterns and statistical significance were unchanged. Synthesis of PGFM in cows fed diets high in LA prepartum was greater postpartum possibly due to the in creased intake of the direct precursors to the 2 series prostaglandins. Reproductive Measurements Size of the previous pregnant horn at 21 and 28 DIM did not differ between treatments or parities, nor di d the change in uterine horn size from 21 to 28 DIM (Table 3.5). At 21 DIM, mulitparous cows fed CSLCFA prepartum had more uterine tonus than multiparous cows not fed CSLCFA prepartum, however this was not evident in primiparous animals (CSLCFA prepartu m vs. no CSLCFA prepartum by parity

PAGE 80

67 interaction; P = 0.050). This pattern did not carry over to measurements taken at 28 DIM. There were no differences between treatmen ts or parities in the size of the cervical os at 21 DIM (3.38 cm). However at 28 DIM, multiparous cows fed CSLCFA prepartum tended to have a smaller cervical os than those not fed CSLCFA prepartum (2.97 vs. 3.30 cm) whereas this did not occur in primiparous cows fed CSLCFA prepartum versus those not fed CSLCFA (3.12 vs. 2.89 cm) (parit y by CSLCFA prepartum vs. no CSLCFA prepartum interaction, P = 0.089). There were no differences between treatments or parities in whether the cervix was open or closed at 21 a nd 28 DIM (Table 3.5). At 21 DIM, cervical color was red only for some pr imiparous cows fed CSLCFA prepartum. Therefore the CSLCFA prepar tum vs. no CSLCFA prepartum by parity interaction was significant (P < 0.001). Cervical color at 28 DI M was not affected by treatment or parity (Table 3.5). Clinical endometritis as described by a purulent or foul discharge after 20 d postpartum or a mucopurulent discharge afte r 26 d postpartum was associated with a reduction of pregnancy rates (LeBlanc et al., 2002). In addition, abnormal vaginal discharge has been correlated with a delay in the first pos tpartum ovulation (Opsomer et al., 2000). Furthermore, if the first ovulation occurred in the presence of a uterus with heavy contamination, it led to prolonged luteal phases which was also associated with lower fertility (Opsomer et al., 2000). Primiparous cows had a greater (more purulent discharge) average score for vaginal disc harge amount at 21 DIM in comparison to multiparous cows (2.32 vs. 1.77; P = 0.034) however at 28 DIM there were no differences between treatments or parities (Table 3.6). Primiparous cows had a greater average score for vaginal discharge quality (g reater evidence of inf ection) at 21 DIM in

PAGE 81

68 comparison to multiparous cows (3.37 vs. 1.42; P < 0.001), however at 28 DIM differences among treatments or between pari ties were not detected (Table 3.6). Primiparous cows had less class 1 (P = 0.048) and class 2 follicles (P = 0.034) at 21 DIM in comparison to multiparous cows (Table 3.7), but the number of class 3 follicles tended to be greater at 28 DIM (P = 0.099) in primiparous compared to multiparous cows (Table 3.7). The number of class 1 (8.0 vs 15.5; P = 0.09) and class 2 (0.7 vs. 2.8; P = 0.10) follicles at 21 DIM was less in multiparou s cows fed CSLCFA prepartum compared to cows not fed CSLCFA prepartum (Table 3.7 ). However, there were no such changes in primiparous cows thus accounting for th e interaction. These changes in follicle dynamics at 21 DIM reflects stimulation in th e number of class 3 follicles for multiparous cows due to fat feeding prepartum. Number of class 3 follicles at 21 DIM were increased in multiparous cows fed CSLCFA prepartum (2.3 vs. 1.4; P = 0.06) with no differences among diets for the number of class 3 follicles of primiparous cows. This early stimulus in pre-ovulatory follicles (cla ss 3) due to fat feeding prep artum was eliminated among the other groups by 28 DIM with primiparous cows tending to have greater overall number of class 3 follicles (2.2 vs. 1.6; P = 0.10). At 21 DIM, the number of CL tended to be fewer (0.4 vs. 0.9) and smaller (16.9 vs. 25.2 mm) for cows fed CSLCFA starting at 1 DIM compared to those not fed CSLCFA (Table 3.7). The number and si ze of CL present on the ovaries at 28 DIM did not differ between treatments or parities. Stimulation of ovarian follicle developmen t in cows fed supplemental fat often has been reported (Staples et al., 1998). Fa t supplementation for 21 d prepartum did not affect follicle dynamics in cows fed isoene rgetic diets containing 1.7% supplemental fat (prilled long chain FA) (Frajb lat and Butler, 2003). However supplemental fat prepartum

PAGE 82

69 resulted in greater pregnancy rates (86 vs 58%). In contrast, workers in Missouri (Oelrichs et al., 2004) recently reported th at cows fed soybeans beginning either prepartum or at calving had fewer small (< 5 mm) follicles and tended to have more medium size (6 to 9 mm) follicles than controls during the first synchronized estrous cycle. During the first estrous synchr onization in the pres ent study, there was no difference in number of class 2 follicles (6 to 9 mm), class 3 follicles (> 10 mm), number of CL, or size of CL present as determ ined by ultrasound on the day that GnRH was injected (d 0), nor on d 2, 4, 6, or 7 following GnRH (Table 3.8). Such changes were evident in the present stu dy through 21 and 28 d postpartum. However, changes thereafter during the synchronization period we re not detected. Fo r example, during the same time frame, the number of class 1 follicles (2 to 5 mm) was fewer for cows fed CSLCFA prepartum compared to cows fed CS LCFA beginning in the postpartum period (10.4 vs. 14.8 averaged across the five m easurement periods; P = 0.008). This may reflect a greater turnover of follicles in this group. Improvement in conception rates when fat is supplemented in the diet is reported often (Staples et al., 1998). No change in conception rates due to fat feeding also is regularly reported (Holter et al ., 1992; Oelrichs et al., 2004). In contrast, a few studies have reported a decrease in conception rate s of cows fed supplemental fat (Erickson et al., 1992; Sklan et al., 1994), al though it may have been due to greater milk production and a more negative energy balance which ha ve been strongly linked to decreased fertility in dairy cattle. In this study, first service conception rates of cows that responded to synchronized ovulation were 27% (3/11), 40% (4/10), 70% (7/10), and 63% (7/11) for cows fed no CSLCFA, CSLCFA prepartum, CS LCFA beginning at 1 DIM, and CSLCFA

PAGE 83

70 beginning at 28 DIM, respectively. Cows fe d CSLCFA tended to have greater first service conception rates compar ed to cows not fed CSLCFA (58 vs. 27%, respectively, P < 0.10). The number of cows per treatment in this study were too low to have a lot of confidence in the conception rate results however a 31% increase is noteworthy. Although improvement in first service concepti on rate was not due to differences in follicular dynamics during the first synchronized estrous, IGF-I acts synergistically with LH to promote follicular development (Lucy, 2001). Greater concentrations of plasma IGF-I around the time of breeding in multip arous cows fed CSLCFA (Figure 3.9) may have contributed to increased first service conception rates. Hepatic Measurements In the present study, mean hepatic TAG concentrations tended to be greater for cows fed CSLCFA beginning prepartum in comparison to cows not fed CSLCFA prepartum (16.7 vs. 10.4% of dry liver we ight; P = 0.080; Table 3.9). Hepatic TAG concentrations in cows fed CSLCFA prepar tum and control diets peaked at 14 DIM, however, cows fed CSLCFA beginning at 1 DIM did not decrease at 28 DIM but remained elevated (treatment by day interaction; P = 0.04; Figure 3.15). At 14 DIM the TAG concentration of cows fed CSLCFA prep artum tended to be greater than that of cows not fed CSLCFA prepartum (P = 0.084; Figure 3.15). Plasma NEFA concentrations were greater in cows fed CS LCFA prepartum than cows not fed CSLCFA prepartum in early lactation, which ma y account for the differences in TAG accumulation. In contrast, Illinois workers (G rum et al., 1996) reported decreased liver TAG concentrations at 1 DIM and a tendency for decreased plasma NEFA concentrations early postpartum in cows fed 6.5% of DM as fa t for 50 d prepartum. They also reported a

PAGE 84

71 positive correlation between concentrations of plasma NEFA at 3 d prepartum and the concentration of TG in the liver at 1 d postp artum. However, a la ter study (Douglas et al., 2004) in which cows were fed a moderate non-fi ber carbohydrate ( NFC ) control diet, a low NFC diet supplemented with 4% fa t prepartum and 2% fat postpartum, or a moderate NFC fat-supplemented diet beginni ng at 60 d prepartum revealed no treatment differences in DMI, milk production, plasma concentration of NEFA, or total hepatic lipid or TAG at 1 DIM. Additionally, recent research conducted at the University of Florida found no difference in hepatic TAG accu mulation in cows fed control, 225 g/d of Ca salts of conjugated linoleic acid ( CLA ), or 225 g/d of Ca salts of trans C18:1 beginning at 28 d prepartum despite differences in plasma NEFA concentrations at 1 wk postpartum (Selberg et al., 2004). While othe rs have reported a negative effect of elevated hepatic TAG on day to first estrus (Jorritsma et al., 2000) or to first ovulation (Marr et al., 2002), days to fi rst ovulation in this study was not greater for the treatment group with elevated hepatic TAG c oncentration (CSLCFA prepartum). Neither treatment nor parity affected he patic IGF-I or IGF-II mRNA expression (Table 3.9; Figures 3.17 and 3.18). The liver is the primary source of circulating IGF-I and decreased hepatic IGF-I mRNA expres sion results in decreased circulating concentrations of IGF-I in cattle (Wang et al ., 2003). In this study, the additional hepatic lipid present postpartum in cows fed CSLCFA prepartum was not detrimental to IGF-I or IGF-II mRNA expression. Cows fed CSLCFA beginning prepartum tended to have less IGFBP-2 mRNA expression in comparison to cows not fed CSLCFA prepartum (0.078 vs. 0.121 arbitrary units/18S; P = 0.097; Figur e 3.19). Steady state expression of IGF-II mRNA expression, which is an indirect measur ement of synthesis, was not affected by

PAGE 85

72 treatment and steady state IGFBP-2 expr ession was lower in cows fed CSLCFA prepartum, indirectly implying that circul ating IGF-II in cows fed CSLCFA prepartum was increased. Recently, University of Flor ida workers reported no treatment differences in hepatic IGF-I mRNA expression in cows fed a control diet, 225 g/d of calcium salts of CLA, or 225 g/d of trans C18:1 during early lactation (Sel berg et al., 2003). However, IGF-II mRNA and IGFBP-2 abundan ce were greater for cows fed trans C18:1 than for cows fed the control diet or CLA. Expr ession of mRNA of IG F-1 (Figure 3.17), IGF-II (Figure 3.18), and IGFBP-2 (Figure 3.19) di d not differ among biopsy days nor were treatment by day interactions detected to be significant for th ese three dependent variables. Measurements of Immune Status Immune reactions have been shown to be modulated by the diet, including the PUFA composition of the diet (Calder et al., 2002). Mechan isms involved in regulation are not yet understood, but evidence exists th at the poly unsaturated FA composition of the diet influences cellular communicati on and activation through the synthesis of prostaglandins, tumor necrosis factor, and interferon(Calder et al., 2002). Linoleic acid can be converted to arachidonic aci d, the precursor for prostaglandin E2 and leukotriene B4 which are pro-inflammatory mediators. Lessard et al. (2004) evaluated cellular immune functions of dairy cows fed supplemental fat during the transition period. Cows were fed a diet containing 2.7% Ca salt of palm oil, 5.9% flaxseed (n-3 FA), or 9.4% micronized soybeans (n-6 FA) from 6 wk prepartum to calving followed by diets containing and 4.7% Ca salt of palm oil, 9.7% flaxseed, or 20.3% micronized soybeans from calving to 6 wk postpartum. The authors concluded that cellular immune functions were modulated around parturition; ho wever feeding diets rich in n-3 or n-6 FA

PAGE 86

73 did not have a major impact on cellular imm une function. In the present study, incidence of disease (mastitis, metritis, or RFM) duri ng the first 10 DIM was lower (P < 0.05) for cows fed CSLCFA prepartum (8%, 1/12) vers us those not fed CSLCFA prepartum (35% [8/23] for the no CSLCFA treatment and 58% [7/12] for the CSLCFA at 1 DIM treatment) which could have had benefici al effects on milk production and reproduction. In response to immunological stress, the liver will produ ce the acute phase proteins ceruloplasmin, fibrinogen, and haptoglobin (Bau mann and Gauldie, 1994). It is unclear whether increased concentrations of acute phase proteins in l actating dairy cattle is due to greater stress (indicating an adverse stat e) or due to a greater immune response (indicating a healthier state). Fibrinogen is involved with blood clotting and the formation of the fibrin matrix for tissue repa ir. Increased fibrinogen concentrations are detected during internal hemorrhage or tissu e damage. Normal values in cattle range from 100 to 600 mg/dl (The Merck Veterinary Manual, 1997). In this study, the range was 26 to 402 mg/dl. The concentrations rose plateaued, then decr eased during the first 4 wk postpartum. Mean plasma concentr ations of fibrinogen during the first 27 d postpartum tended to be greater for control co ws than for cows fed CSLCFA beginning at 1 DIM (123.8 vs. 100.2 mg/dl; P = 0.07; Table 3.10). Primiparous cows had greater fibrinogen concentrations when CSLCFA was withheld prepartum than when fed prepartum (114.4 vs. 70.5 mg/dl); however fibri nogen concentrations were not different for multiparous cows regardless of when CSLCFA feeding was initiated (no CSLCFA prepartum vs. CSLCFA prepar tum by parity interaction; P = 0.029; Table 3.10). Haptoglobin is responsible for binding iron, a limiting nutrient in bacterial growth. Because haptoglobin concentrations are nor mally undetectable in bovine blood unless

PAGE 87

74 there is tissue damage, it can be a good indica tor of the immunological stress response to parturition. In this study, the range was 0 to 329 mg of HbB/ 100 ml (the amount of hemoglobin bound by haptoglobin/ 1 00 ml of plasma). Mean plasma concentrations of haptoglobin during the first 27 d postpartu m were greater for primiparous than multiparous cows (31.9 vs. 17.0 mg of HbB/ 100 ml of plasma; P = 0.043; Table 3.10), indicating that parturition caused a greater im mune response in primiparous cows. Using heterogeneity of regression, the cubic pa ttern of plasma haptoglobin over time was affected by feeding CSLCFA and parity (T able A-5). Primiparous cows not fed CSLCFA prepartum had greater initial plasma concentrations of haptoglobin than cows fed CSLCFA prepartum and declined at a fa ster rate so that both treatment groups reached their nadir at 23 DIM (Figure 3.20A). Multiparous cows fed CSLCFA prepartum had very low initial concentrations of haptoglobin, rose to peak concentrations at 7 DIM and reached their nadir at 23 DIM. Multiparous cows not fed CSLCFA prepartum had higher initial c oncentrations and steadily d eclined through 27 DIM (Figure 3.20B). The trends over time indicate cows fe d CSLCFA prepartum had less of an acute phase protein response to parturition than cows not fed CSLCFA prepartum, although it is unclear why haptoglobin concentrations in multiparous cows fed CSLCFA prepartum did not peak until 7 DIM. Ceruloplasmin is involved with copper tran sport to tissues util izing its antioxidant properties and concentrations wi ll increase due to an inflammatory response of the cow. Normal values in cattle range from 16.8 to 34.2 mg/dl (The Merck Veterinary Manual, 1997). In this study, the range was from 9.0 to 51.0 mg/dl, suggestive of an acute inflammatory response to parturition. Mean plasma concentrations of ceruloplasmin

PAGE 88

75 were greater for primiparous than for multiparous cows during the first 27 d postpartum (24.4 vs. 20.7 mg/dl; P = 0.007) indicating that parturition caused a greater immune response in primiparous cows. The pattern over time was not different among treatments or between parities (Figure 3.21). Elevations in plasma concentrations of total biliruben are due to severe hepatic damage or extrahepatic obstruction, and followi ng pregnancy. Normal values in cattle range from 0 to 0.5 mg/dl (The Merck Veterina ry Manual, 1997). In this study, the range was 0.0 to 1.3 mg/dl. Mean plasma concentrat ions of total bilirub en were greater for cows fed CSLCFA prepartum than for co ws not fed CSLCFA prepartum (0.36 vs. 0.23 mg/dl; P = 0.04), although values were still wi thin the normal range. An effect of week postpartum was detected (P < 0.001; Figure 3.22). Elevations in plasma concentrations of ALT are due to hepatic disease although it is not a good indicator in cattle because of its low activity and it is not liver specific (The Merck Veterinary Manual, 1997). Mean plas ma concentrations of ALT did not differ among treatments or between parities (21.6 IU/L; Table 3.10), however an effect of week postpartum was detected (P = 0.003; Figure 3.23). Elevations in plasma concentrations of ALK are due to liver lesions or bile duct obstruction, however in cattle th ere is a very wide range of normal activity between animals (The Merck Veterinary Manual, 1997). Mean plasma concentrations of ALK were lower for multiparous cows in comparison to primiparous cows (31.7 vs. 44.1 U/L; Table 3.10). An effect of week postpartu m was detected (P < 0.001; Figure 3.24). Concentrations of AST are pr esent in most tissues and it is not liver specific. Damage to the liver and the postparturient peri od in cattle cause leak age of large amounts

PAGE 89

76 of AST into blood (The Merck Veterinary Ma nual, 1997). Mean plasma concentrations of AST did not differ among treatments or between parities ( 90.5 IU/L; Table 3.10), however an effect of week postpartum wa s detected (P = 0.01; Figure 3.25). Plasma concentrations of GGT are of hepa tic origin and can be a good indicator of liver and bile duct damage in cattle (The Me rck Veterinary Manual, 1997). Mean plasma concentrations of GGT did not differ among treatments or between parities (30.0 U/L; Table 3.10), however an effect of week postpar tum was detected (P = 0.01; Figure 3.25). At wk 4 postpartum cows fed CSLCFA prepar tum had greater plasma concentrations of GGT than cows not fed CSLCFA prepartum (P = 0.03; Figure 3.26). Plasma concentrations of albumin d ecrease due to the failure of hepatic parenchymal synthesis. In cattle, only 5% of serum albumin levels are synthesized per day, therefore it takes time to see hepatic da mage (The Merck Veterinary Manual, 1997). Concentration of plasma albumin increased slightly with wk postpartum (P = 0.003; Figure 3.27). Initiation of CSLCFA supplementation at calving stimulated albumin concentrations in plasma of primiparous cows but reduced it in multiparous cows (no CSLCFA vs. CSLCFA 1 DIM by parity interaction; P = 0.018; Table 3.10). Parity Effects As expected, primiparous cows weighed less, produced less milk, and tended to have less SCC in the milk than multiparous cows. In addition, primiparous cows had greater circulating glucose and tended to have lower circulating BUN then multiparous cows. Parturition, removal of the calf, and th e milking process is probably more stressful on first calf heifers than on multiparous cows. Indicative of a higher stress level, heifers had a greater immune response soon after part urition than cows as indicated by greater circulating concentration of the acute phase proteins, haptoglobin and ceruloplasmin, and

PAGE 90

77 the liver enzyme ALK. At 21 DIM, primip arous cows had more vaginal discharge, a more purulent vaginal discharge, and fewer class 1 and class 2 follicles and tended to have more class 3 follicles at 28 DIM than multiparous cows. Conclusion Holstein cows began consuming a diet at 0 or 2% of dietary DM as CSLCFA at ~28 d prepartum. Cows fed the 0% CSLCFA diet either remained on a CSLCFA-free diet at parturition or were shifted to a 2% CSLCFA diet at ei ther 1 or 28 DIM and remained on said diet until 100 DIM. Animals fed CSLCFA in the prepartum period continued to receive CSLCFA throughout the lactation pe riod. Cows fed CSLCFA beginning in the perpartum peri od tended to produce more milk. This milk increase was accompanied by a tendency for elevated concentr ations of TG in the liver at 14 DIM, a tendency for lower expression of hepatic IG FBP-2 mRNA, and elevated concentrations of plasma biliruben compared to cows not receiving CSLCFA prepartum. Multiparous cows appeared to benefit more from suppl ementation with CSLCFA than primiparous cows in that multiparous cows not fed CS LCFA at any time during the study had or tended to have lower concentr ations of milk protein, l onger and greater loss of BW, greater concentrations of plasma BHBA, and lower concentrations of plasma glucose, leptin, and IGF-I. In addition, multiparous cows fed CSLCFA prepartum tended to have fewer small and medium size but more larger size follicles ( 10 mm in size), more uterine tone at 21 DIM a smaller cervical os at 28 DIM, and a slower decrease in plasma concentrations of PGFM the first 10 d postpartum than multiparous cows not fed CSLCFA prepartum. Multiparous cows fed CS LCFA had greater concentrations of IGFI at the time of AI and all cows fed CSLCFA tended to have greater conception rate at first service regardless of day of initiation of CSLCFA supplementation.

PAGE 91

78 Table 3.1: Ingredient and chemical com position of diets fed to nonlactating cows. Control CSLCFA1 Ingredient, % of DM Corn silage 50 50 Bermudagrass hay 10 10 Ground corn 16.9 14.4 Citrus pulp 5.0 5.1 Soybean meal 11.8 12.4 MegalacR 0.0 2.0 Mineral mix 2 6.2 6.2 Trace mineral salt 3 0.009 0.009 Chemical DM% 39.8 39.8 NEL, Mcal/kg of DM 1.58 1.61 CP, % of DM 15.1 14.3 Ether extract, % of DM 3.1 4.4 NDF, % of DM 36.2 36.7 ADF, % of DM 21.3 21.6 Ca, % of DM 2.2 2.1 P, % of DM 0.33 0.32 K, % of DM 1.1 1.1 Mg, % of DM 0.35 0.33 Na, % of DM 0.24 0.25 Cl, % of DM 1.04 0.92 S, % of DM 0.26 0.26 Mn, mg/kg of DM 34 33 Cu, mg/kg of DM 12 14 Zn, mg/kg of DM 42 47 Fe, mg/kg of DM 257 249 Mo, mg/kg of DM 1.4 1.6 1 MegalacR, Church & Dwight Co. Inc., Princeton, NJ. 2 Mineral Mix contained 22.8% CP, 22.9% Ca 0.20% P, 0.2% K, 2.8% Mg, 0.7% Na, 2.4% S, 8% Cl, 147 mg/kg of Mn, 27 mg/kg of Fe, 112 mg/kg of Cu, 95 mg/kg of Zn, 7 mg/kg of Se, 8 mg/kg of I, 11 mg/kg of Co, 268,130 IU of vitamin A/kg, 40,000 IU of vitamin D/kg, and 1129 IU of vitamin E/kg (DM basis). 3 Minimum concentrations of 40% Na 55% Cl, 0.25% Mn, 0.2% Fe, 0.033% Cu, 0.007% I, 0.005% Zn, and 0.0025% Co (DM basis).

PAGE 92

79 Table 3.2: Ingredient and chemical com position of diets fed to lactating cows. Control CSLCFA1 Ingredient, % of DM Corn silage 37.5 37.5 Alfalfa hay 10 10 Ground corn 24.2 21.9 Citrus pulp 5.0 5.0 Cottonseed hulls 2.5 2.5 Soybean meal 9.1 9.4 Soy Plus 2 6.8 6.8 Bio Phos 3 0.4 0.4 MegalacR 0.0 2.0 Mineral mix 4 4.5 4.5 Chemical DM% 46.2 46.3 NEL, Mcal/kg of DM 1.67 1.71 CP, % of DM 16.9 16.7 Ether extract, % of DM 3.1 4.7 NDF, % of DM 30.9 30.2 ADF, % of DM 18.9 18.8 Ca, % of DM 1.15 1.51 P, % of DM 0.49 0.46 K, % of DM 1.27 1.27 Mg, % of DM 0.29 0.31 Na, % of DM 0.44 0.53 Cl, % of DM 0.34 0.39 S, % of DM 0.23 0.24 Mn, mg/kg of DM 71 86 Cu, mg/kg of DM 23 23 Zn, mg/kg of DM 88 106 Fe, mg/kg of DM 236 227 Mo, mg/kg of DM 2.1 1.6 1 MegalacR, Church & Dwight Co. Inc., Princeton, NJ. 2 West Central Soy, Ralston, IA. 3 IMC-Agrico, Bannockburn, IL. 4 Mineral Mix contained 26.4% CP, 10.2% Ca, 0.90% P, 3.1% Mg, 1.5 % S, 5.1% K, 8.6 % Na, 11698 mg/kg of Zn, 512 mg/kg of Cu, 339 mg/kg of Fe,2231 mg/kg of Mn, 31 mg/kg of Co, 26 mg/kg of I, 7.9 mg/kg of Se, 147,756 IU of vitamin A/kg, and 787 IU of vitamin E/kg (DM basis).

PAGE 93

80Table 3.3. Milk yield, milk composition, post partum body weight, and postpartum body conditi on score of Holstein cows fed diets without calcium salts of long chain fatty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum, at 1 day in milk (DIM) or at 28 DIM. Treatments Orthogonal contrasts 1, P = Measure No CSLCFA CSLCFA prepartum CSLCFA 1 DIM CSLCFA 28 DIM SE A B C Parity D E F P2 M3 P M P M P M Milk yield, kg/d 36.0 41.0 39.5 44.8 35.4 38.3 34.7 39.9 3.1 0.924 0.059 0.896 0.041 0.933 0.816 0.710 3.5% FCM, kg/d 36.1 39.7 40.2 42.5 34.7 37.3 35.4 38.5 3.3 0.947 0.087 0.779 0.215 0.861 0.920 0.941 Milk fat, % 3.45 3.28 3.55 3.33 3.53 3.41 3.57 3.42 0.13 0.487 0.789 0.871 0.195 0.977 0.782 0.949 Milk protein, % 2.63 2.47 2.58 2.69 2.56 2.70 2.75 2.71 0.56 0.102 0.525 0.241 0.846 0.091 0.679 0.261 Milk fat yield, kg/d 1.27 1.36 1.43 1.44 1.21 1.28 1.26 1.34 0.09 0.912 0.114 0.659 0.451 0.865 0.783 0.967 Milk protein yield, kg/d 1.03 1.06 1.06 1.25 0.92 1.06 1.01 1.10 0.06 0.756 0.080 0.439 0.067 0.487 0.649 0.797 Milk SCC, x 1000/ml 407 750 463 466 264 700 282 428 121 0.324 0.760 0.465 0.067 0.614 0.400 0.407 BW, kg 530 644 539 621 518 646 533 664 28 0.991 0.676 0.555 < 0.001 0.992 0.325 0.957 BCS 3.06 2.77 2.88 3.10 3.02 3.31 3.13 2.95 0.19 0.363 0.501 0.534 0.937 0.220 0.626 0.231 1 Orthogonal contrast of means were the following: A = No CSLCFA vs. CSLCFA, B = CSLCFA prepartum vs. (CSLCFA 1 DIM plus CSLCFA 28 DIM), C = CSLCFA 1 DIM vs. CSLCFA 28 DIM, D = contrast A by parity, E = contrast B by parity, and F = contrast C by parity. 2 Primiparous cows. 3 Multiparous cows.

PAGE 94

81Table 3.4. Concentration of plasma hormones and metabolites and day of first ovulation of Holstein cows fed diets without calci um salts of long chain fatty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA star ting prepartum, at 1 day in milk (DIM) or at 28 DIM. Treatments Orthogonal contrasts 1, P = Measure No CSLCFA CSLCFA prepartum CSLCFA 1 DIM CSLCFA 28 DIM SE A B C Parity D E F P2 M3 P M P M P M BHBA, mg/dl 10.7 14.2 10.4 9.7 10.2 6.7 8.4 6.9 1.6 0.007 0.146 0.629 0.635 0.044 0.501 0.520 NEFA, meq/L 384 450 432 480 400 241 289 245 56 0.149 0.002 0.348 0.588 0.217 0.130 0.316 Glucose, mg/dl 65.5 55.2 65.2 60.4 68.7 64.8 67.8 66.5 2.1 0.004 0.025 0.846 0.001 0.054 0.553 0.533 Blood urea nitrogen, mg/dl 10.2 11.5 10.4 15.3 9.8 10.9 10.6 11.0 1.3 0.682 0.053 0.744 0.055 0.706 0.081 0.808 Leptin, ng/ml 3.06 1.65 2.83 2.62 1.91 1.99 1.70 2.57 0.39 0.846 0.156 0.729 0.669 0.078 0.465 0.470 IGF-I, ng/ml 77.1 54.3 62.2 60.6 64.3 69.9 66.2 74.1 7.5 0.953 0.261 0.683 0.641 0.040 0.487 0.885 Insulin, ng/ml 0.62 0.54 0.65 0.52 0.71 0.75 0.74 0.63 0.09 0.236 0.102 0.608 0.320 0.975 0.536 0.449 Accumulated progesterone, 1 to 77 DIM, ng/ml 22.0 27.4 24.0 24.0 21.9 26.2 26.5 23.3 4.0 0.948 0.884 0.779 0.555 0.516 0.906 0.602 Accumulated progesterone, 1 to 46 DIM, ng/ml 7.3 11.1 8.7 10.9 7.9 9.7 9.1 8.0 2.4 0.942 0.806 0.930 0.467 0.606 0.931 0.847 DIM at first ovulation 27.8 26.3 29.7 27.8 32.2 27.5 22.8 24.6 4.0 0.910 0.560 0.125 0.587 0.987 0.955 0.417 1 Orthogonal contrast of means were the following: A = No CSLCFA vs. CSLCFA, B = CSLCFA prepartum vs. (CSLCFA 1 DIM plus CSLCFA 28 DIM), C = CSLCFA 1 DIM vs. CSLCFA 28 DIM, D = contrast A by parity, E = contrast B by parity, and F = contrast C by parity. 2 Primiparous cows. 3 Multiparous cows.

PAGE 95

82Table 3.5. Concentration of plasma PGF2 metabolite (PGFM) the first 14 DIM and the size and characteristic s of the uterus and cervix of Holstein cows fed diets without calcium salts of long chain fatty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum, or at 1 day in milk (DIM). Treatments Orthogonal contrasts 1, P = Measure No CSLCFA CSLCFA prepartum CSLCFA 1 DIM SE A B Parity C D P2 M3 P M P M PGFM, pg/ml 1004 1186 1443 1412 1536 955 244 0.261 0.722 0.418 0.509 0.111 Previous pregnant horn size at 21 DIM, cm 2.43 2.52 2.38 2.62 2.55 3.95 0.18 0.500 0.133 0.114 0.984 0.376 Previous pregnant horn size at 28 DIM, cm 2.24 2.60 2.50 2.35 2.42 2.59 0.20 0.841 0.655 0.452 0.256 0.613 Change in uterine horn size from 21 to 28 DIM, cm -0.20 0.08 0.12 -0.27 -0.13 -0.36 0.20 0.683 0.357 0.456 0.269 0.199 Uterine tonus4 at 21 DIM 1.80 1.77 1.50 2.17 2.17 1.50 0.28 0.923 0.855 0.964 0.050 0.237 Uterine tonus4 at 28 DIM 1.70 1.69 1.33 2.00 1.67 1.83 0.26 0.815 0.832 0.209 0.227 0.731 Size of cervical os at 21 DIM, cm 3.20 3.59 3.35 3.54 3.25 3.38 0.19 0.610 0.663 0.140 0.852 0.480 Size of cervical os at 28 DIM, cm 3.12 3.23 3.12 2.97 2.67 3.38 0.18 0.727 0.387 0.136 0.089 0.086 Cervix closed (1) or open (2) at 21 DIM 1.20 1.00 1.00 1.00 1.00 1.00 0.07 0.460 0.164 0.277 0.460 0.164 Cervix closed (1) or open (2) at 28 DIM 1.10 1.00 1.00 1.00 1.00 1.00 0.05 0.622 0.350 0.467 0.622 0.350 Cervical color, pink (1) or red (2) at 21 DIM 1.00 1.00 1.50 1.00 1.00 1.00 0.07 < 0.001 1.00 0.007 < 0.001 1.00 Cervical color, pink (1) or red (2) at 28 DIM 1.00 1.08 1.17 1.00 1.00 1.00 0.08 0.367 0.606 0.639 0.152 0.606 1 Orthogonal contrast of means were the following: A = (No CSLCFA plus CSLCFA 1 DIM) vs. CSLCFA prepartum, B = No CSLCFA vs. CSL CFA 1 DIM, C = contrast A by parity, and D = contrast B by parity. 2 Primiparous cows. 3 Multiparous cows. 4 Score of 1 = no tonus, 2 = moderate tonus, and 3 = tonus.

PAGE 96

83Table 3.6. Vaginal observations at 21 and 28 DIM of Holstein cows fed diets without calcium salts of long chain fatty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA star ting prepartum, or at 1 day in milk (DIM). Treatments Orthogonal contrasts 1, P = Measure No CSLCFA CSLCFA prepartum CSLCFA 1 DIM SE A B Parity C D P2 M3 P M P M Vaginal discharge amount4 at 21 DIM 2.30 2.15 2.33 1.50 2.33 1.67 0.30 0.481 0.441 0.034 0.445 0.377 Vaginal discharge amount4 at 28 DIM 2.40 1.46 1.83 2.00 2.17 1.83 0.33 0.872 0.828 0.182 0.189 0.344 Vaginal discharge quality5 at 21 DIM 3.60 1.92 2.67 1.00 3.83 1.33 0.60 0.133 0.759 < 0.001 0.702 0.479 Vaginal discharge quality5 at 28 DIM 2.50 1.08 1.50 1.67 3.33 2.00 0.69 0.313 0.193 0.137 0.228 0.946 1 Orthogonal contrast of means were the following: A = (No CSLCFA plus CSLCFA 1 DIM) vs. CSLCFA prepartum, B = No CSLCFA vs. CSL CFA 1 DIM, C = contrast A by parity, and D = contrast B by parity. 2 Primiparous cows. 3 Multiparous cows. 4 Score of 1 = none, 2 = trace, 3 = slight, 4 = moderate, or 5 = copious. 5 Score of 0 = none, 1 = clear, 2 = cloudy, 3 = mucous with pus, 4 = mucopurulent, or 5 = purulent.

PAGE 97

84Table 3.7. The number and size1 of ovarian structures at 21 and 28 DIM of Holstein cows fed diets without calcium salts of long chain fatty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartu m, or at 1 day in milk (DIM). Treatments Orthogonal contrasts 2, P = Measure No CSLCFA CSLCFA prepartum CSLCFA 1 DIM SE A B Parity C D P3 M4 P M P M Number of class 1 follicles at 21 DIM 8.4 12.8 9.0 8.0 10.7 18.3 2.2 0.050 0.074 0.048 0.087 0.451 Number of class 2 follicles at 21 DIM 1.1 2.8 0.8 0.7 0.5 2.8 0.7 0.111 0.695 0.034 0.100 0.627 Number of class 3 follicles at 21 DIM 1.4 1.5 1.0 2.3 1.8 1.3 0.4 0.718 0.780 0.358 0.057 0.437 Number of CL5 at 21 DIM 0.7 1.2 0.8 1.0 0.3 0.5 0.3 0.442 0.080 0.278 0.756 0.555 Size of CL at 21 DIM, mm 22.9 27.5 23.0 29.0 12.3 21.5 4.6 0.256 0.084 0.103 0.915 0.616 Number of class 1 follicles at 28 DIM 14.0 14.3 10.5 9.7 12.0 16.3 2.3 0.058 0.995 0.504 0.454 0.365 Number of class 2 follicles at 28 DIM 2.0 2.5 1.3 1.5 0.7 1.3 0.7 0.723 0.055 0.411 0.723 0.921 Number of class 3 follicles at 28 DIM 2.1 1.7 2.7 1.8 1.8 1.5 0.4 0.181 0.530 0.099 0.505 0.919 Number of CL at 28 DIM 0.9 0.9 0.7 1.2 0.8 1.2 0.3 0.880 0.746 0.227 0.536 0.571 Size of CL at 28 DIM, mm 27.1 24.3 27.0 34.4 33.2 26.9 5.3 0.581 0.396 0.903 0.295 0.729 1 Class 1 follicles were 2 to 5 mm, class 2 follicles were 6 to 9 mm, and class 3 follicles were 10 mm. 2 Orthogonal contrast of means were the following: A = (No CSLCFA plus CSLCFA 1 DIM) vs. CSLCFA prepartum, B = No CSLCFA vs. CSL CFA 1 DIM, C = contrast A by parity, and D = contrast B by parity. 3 Primiparous cows. 4 Multiparous cows. 5 Corpus luteum.

PAGE 98

85Table 3.8.Ovarian structures present on the ova ries of Holstein cows fed diets without calcium salts of long chain fatty acids (CSLCFA) (no CSLCFA) or diets with CS LCFA starting prepartum, at 1 day in milk (DIM) or at 28 DIM where day equals the days after GnRH injection for estrous synchroni zation (44 3 DIM). Treatments Measure Day No CSLCFA CSLCFA prepartum CSLCFA 1 DIM CSLCFA 28 DIM SE 0 14.0 9.1 15.2 13.4 1.5 2 11.2 11.5 15.3 19.1 1.4 4 12.6 9.9 16.3 13.8 1.4 6 12.3 11.0 15.7 12.1 1.4 Number of class 1 follicles a (2 to 5 mm) 7 12.8 10.5 16.4 10.3 1.4 0 1.5 1.0 3.25 2.1 0.5 2 2.2 3 3.4 2.1 0.5 4 2.3 1.4 1.4 3.2 0.5 6 2.4 2.3 0.9 2.4 0.5 Number of class 2 follicles (6 to 9 mm) 7 1.8 1.9 2 1.6 0.5 0 2.3 1.8 2.5 2.0 0.3 2 2.1 1 1.3 1.1 0.3 4 2.2 2.1 2.3 1.9 0.3 6 2.4 2.3 1.6 1.9 0.3 Number of class 3 follicles ( 10 mm) 7 2.8 2.5 2.0 2.0 0.3 0 1.0 0.7 0.7 1.0 0.2 2 1.0 0.9 0.9 1.2 0.2 4 1.2 1.25 1.2 1.3 0.2 6 1.3 1.3 1.0 1.7 0.2 Number of corpus luteum b 7 1.4 1.5 1.2 1.7 0.2 0 21.3 11.6 13.4 21.8 3.6 2 21.3 16.7 15.4 25.0 3.5 4 25.8 21.0 21.7 26.6 3.5 6 27.3 25.3 20.2 34.6 3.5 Size of corpus luteum c (mm) 7 28.7 29.0 25.5 29.6 3.5 a CSLCFA prepartum vs. CSLCFA postpartum, P = 0.008. b CSLCFA 1 DIM vs. CSLCFA 28 DIM, P = 0.099. c CSLCFA 1 DIM vs. CSLCFA 28 DIM, P = 0.092.

PAGE 99

86Table 3.9. Concentrations of hepatic triacy lglycerol (TAG), and hepatic IGF-I, IGF-II, and IGF bindi ng protein (BP) -2 mRNA lev els of Holstein cows fed diets without calc ium salts of long chain fatty acids (CSLCF A) (no CSLCFA) or diets with CSLCFA starting prepartum, or at 1 day in milk (DIM). Treatments Orthogonal contrasts 1, P = Measure No CSLCFA CSLCFA prepartum CSLCFA 1 DIM SE A B Parity C D P2 M3 P M P M TAG, % of wet weight 2.8 3.0 5.3 6.0 2.7 5.6 1.3 0.102 0.317 0.257 0.725 0.266 TAG % of dry weight 8.6 9.8 16.2 17.2 8.3 14.7 3.8 0.080 0.531 0.364 0.693 0.478 IGF-I mRNA, arbitrary units 0.024 0.020 0.013 0.003 0.011 0.048 0.073 0.110 0.550 0.494 0.231 0.110 IGF-II mRNA, arbitrary units 0.292 0.288 0.269 0.224 0.296 0.331 0.043 0.159 0.591 0.892 0.429 0.670 IGFBP-2 mRNA, arbitrary units 0.087 0.136 0.091 0.065 0.149 0.111 0.028 0.097 0.534 0.837 0.519 0.142 1 Orthogonal contrast of means were the following: A = (No CSLCFA plus CSLCFA 1 DIM) vs. CSLCFA prepartum, B = No CSLCFA vs. CSL CFA 1 DIM, C = contrast A by parity, and D = contrast B by parity. 2 Primiparous cows. 3 Multiparous cows.

PAGE 100

87Table 3.10. Concentration of plasma acute phas e proteins and liver enzymes of Holstein cows fed diets without calcium salts of long chain fatty acids (CSLCFA) (no CSLCFA) or diets with CS LCFA starting prepartum, or at 1 day in milk (DIM). Treatments Orthogonal contrasts 1, P = Measure No CSLCFA CSLCFA prepartum CSLCFA 1 DIM SE A B Parity C D P2 M3 P M P M Fibrinogen, mg/dl 131.7 115.8 70.5 121.6 97.1 103.2 13.4 0.175 0.070 0.211 0.029 0.353 Haptoglobin, mg HbB/100 ml4 31.6 15.4 27.5 18.2 36.5 17.5 8.4 0.750 0.980 0.043 0.646 0.872 Ceruloplasmin, mg/dl 24.4 21.1 25.1 19.0 23.7 21.9 1.6 0.634 0.974 0.007 0.195 0.675 Total biliruben, mg/dl 0.22 0.26 0.32 0.39 0.23 0.22 0.06 0.040 0.837 0.529 0.658 0.640 Alanine aminotransferase, IU/L 22.5 20.2 24.7 20.7 22.7 18.5 2.7 0.482 0.7749 0.122 0.883 0.719 Alkaline phosphatase, U/L 39.8 32.4 45.8 33.8 46.7 29.0 3.9 0.431 0.646 < 0.001 0.947 0.172 Aspartate aminotransferase, IU/L 111.9 89.6 106.9 114.8 119.6 82.5 19.3 0.576 0.986 0.289 0.294 0.692 Gamma glutamyl transferase, U/L 30.8 25.4 32.8 33.4 26.5 31.0 4.5 0.264 0.880 0.976 0.900 0.265 Albumin, g/dl 2.55 2.73 3.01 2.78 2.95 2.45 0.15 0.104 0.672 0.130 0.772 0.018 1 Orthogonal contrast of means were the following: A = (No CSLCFA plus CSLCFA 1 DIM) vs. CSLCFA prepartum, B = No CSLCFA vs. CSL CFA 1 DIM, C = contrast A by parity, and D = contrast B by parity. 2 Primiparous cows. 3 Multiparous cows. 4 Amount of hemoglobin bound by haptoglobin/100 ml of plasma.

PAGE 101

88 0 10 20 30 40 50 60 1234567891011121314 Week postpartumMilk (kg/d) No CSLCFA CSLCFA prepartum CSLCFA 1 DIM CSLCFA 28 DIM Figure 3.1. Least squares means for milk produc tion of cows fed diets without calcium salts of long chain fatty acids (C SLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum, at 1 day in milk (DIM), or 28 DIM.

PAGE 102

89 450 500 550 600 650 700 750 1234567891011121314 Week PostpartumBody Weight (kg) No CSLCFA CSLCFA prepartum CSLCFA 1 DIM CSLCFA 28 DIM A 450 500 550 600 650 700 750 1234567891011121314 Week PostpartumBody Weight (kg) No CSLCFA CSLCFA prepartum CSLCFA 1 DIM CSLCFA 28 DI M B Figure 3.2. Least squares means for body weight of primiparous (A) and multiparous (B) cows fed diets without calcium salts of long chain fatty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum, at 1 day in milk (DIM), or 28 DIM. There was a treatment x parity x week interaction (P < 0.001).

PAGE 103

90 0 0.5 1 1.5 2 2.5 3 3.5 4 03691214 Week postpartumBody condition score No CSLCFA CSLCFA prepartum CSLCFA 1 DIM CSLCFA 28 DIM Figure 3.3. Least squares means for body condi tion score of cows fed diets without calcium salts of long chain fatty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum, at 1 day in milk (DIM), or 28 DIM. There was a treatment x week inte raction (P = 0.007).

PAGE 104

91 0 5 10 15 20 25 123456789 Week postpartumBHBA (mg/dL) No CSLCFA CSLCFA prepartum CSLCFA 1 DIM CSLCFA 28 DIM* * Figure 3.4. Least squares means for plasma BHBA concentration of cows fed diets without calcium salts of long chain fa tty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum, at 1 day in milk (DIM), or 28 DIM. The asterisks indicate the treatment no CSLCFA is different than CSLCFA 1 DIM and CSLCFA 28 DIM at wk 3, 4, and 5 of lactation and different than the treatment CSLCFA prepartum at wk 5 postpartum (P < 0.05).

PAGE 105

92 0 100 200 300 400 500 600 700 800 900 1000 123456789 Week postpartumNEFA (meq/L) No CSLCFA CSLCFA prepartum CSLCFA 1 DIM CSLCFA 28 DIM* Figure 3.5. Least squares means for plasma NEFA concentration of cows fed diets without calcium salts of long chain fa tty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum, at 1 day in milk (DIM), or 28 DIM. The asterisks indicate that the treatments no CSLCFA and CSLCFA prepartum are different than CSLCFA 1 DIM and CSLCFA 28 DIM at wk 3 and 4 postpartum (P < 0.05).

PAGE 106

93 0 10 20 30 40 50 60 70 80 123456789 Week postpartumGlucose (mg/dll) No CSLCFA CSLCFA prepartum CSLCFA 1 DIM CSLCFA 28 DIM** * Figure 3.6. Least squares means for plasma gl ucose concentration of cows fed diets without calcium salts of long chain fa tty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum, at 1 day in milk (DIM), or 28 DIM. There tended to be a treatment x week interaction (P = 0.10). The asterisks indicate treatment differences at we ek 3, 4, 5, and 6 postpartum (P < 0.05). The treatment no CSLCFA is different than CSLCFA 1 DIM and CSLCFA 28 DIM at wk 3, 4, and 5 postpartum, different than CSLCFA prepartum at wk 5 postpartum, and different than CSLCFA 1 DIM at wk 6 postpartum. The treatment CSLCFA prepartum is differe nt than CSLCFA 28 at wk 3 and different than CSLCFA 1 DIM at wk 6 postpartum.

PAGE 107

94 0 2 4 6 8 10 12 14 16 18 123456789 Week postpartumBUN (mg/dl) No CSLCFA CSLCFA prepartum CSLCFA 1 DIM CSLCFA 28 DIM Figure 3.7. Least squares means for blood urea n itrogen (BUN) concentration of cows fed diets without calcium salts of long ch ain fatty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum, at 1 day in milk (DIM), or 28 DIM. There was a tendency for a treatmen t x week interaction (P = 0.06)

PAGE 108

95 0 0.5 1 1.5 2 2.5 3 3.5 4 123456789 Week postpartumLeptin (ng/ml) No CSLCFA CSLCFA prepartum CSLCFA 1 DIM CSLCFA 28 DI M Figure 3.8. Least squares means for plasma leptin concentration of cows fed diets without calcium salts of long chain fatty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum, at 1 day in milk (DIM), or 28 DIM.

PAGE 109

96 0 10 20 30 40 50 60 70 80 90 1714212835424956637077 Day postpartumIGF-1 (ng/ml) No CSLCFA CSLCFAA 0 10 20 30 40 50 60 70 80 90 1714212835424956637077 Day postpartumIGF-1 (ng/ml) No CSLCFA CSLCFAB Figure 3.9. Regression plot of plasma IGF-1 concentrati on of primiparous (A) and multiparous (B) cows fed diets without cal cium salts of long chain fatty acids (CSLCFA) (no CSLCFA) or cows fed CS LCFA starting prepartum, at 1 day in milk (DIM), or 28 DIM. There was a treatment x parity x day interaction for this contrast (second order polynom ial, P < 0.01). The pooled SE of primiparous cows fed no CSLCFA or CSLCFA was 7.9 and 4.3 ng/ml, respectively. The pooled SE of multiparous cows fed no CSLCFA or CSLCFA was 7.2 and 4.1 ng/ml, respectively.

PAGE 110

97 0 20 40 60 80 100 1714212835424956637077 Day postpartumIGF-1 (ng/ml) CSLCFA prepartum CSLCFA postpartumA 0 20 40 60 80 100 1714212835424956637077 Day postpartumIGF-1 (ng/ml) CSLCFA prepartum CSLCFA postpartumB Figure 3.10. Regression plot of plasma IGF-1 concentrati on of primiparous (A) and multiparous (B) cows fed diets with cal cium salts of long chain fatty acids (CSLCFA) starting prepartum (CSLCFA prepartum) or cows fed CSLCFA beginning at 1 d in milk (DIM) or 28 DIM (CSLCFA postpartum). There was a treatment x parity x day interact ion for this contrast (second order polynomial, P < 0.01). The pooled SE of primiparous cows fed CSLCFA prepartum or CSLCFA postpartum wa s 7.5 and 4.4 ng/ml, respectively. The pooled SE of multiparous cows fe d CSLCFA prepartum or CSLCFA postpartum was 6.7 and 2.6 ng/ml, respectively.

PAGE 111

98 Figure 3.11. Regression plot of plasma insulin concentration of primiparous (A) and multiparous (B) cows fed diets without cal cium salts of long chain fatty acids (CSLCFA) (no CSLCFA), or diets with CSLCFA starting prepartum, at 1 day in milk (DIM), or 28 DIM. There was a treatment x parity x day interaction (second order polynomial, P < 0.01). The pooled SE of primiparous and multiparous cows was 0.05 and 0.05 ng/ml, respectively. 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 151015202530354045505560657075 Day postpartumInsulin (ng/ml) No CSLCFA CSLCFA prepartum CSLCFA 1 DIM CSLCFA 28 DIMA 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 151015202530354045505560657075 Day postpartumInsulin (ng/ml) No CSLCFA CSLCFA prepartum CSLCFA 1 DIM CSLCFA 28 DIMB

PAGE 112

99 0 10 20 30 40 50 60 70 1714212835424956637077 Day postpartumAccumulated progesterone (pg/ml) No CSLCFA CSLCFA A 0 10 20 30 40 50 60 70 1714212835424956637077 Day postpartumAccumulated progesterone (pg/ml) No CSLCFA CSLCFA B Figure 3.12. Regression plot of accumulated plasma progesterone concentration from d 1 to 77 postpartum of primiparous (A) and multiparous (B) cows fed diets without calcium salts of long chain fa tty acids (CSLCFA) (no CSLCFA) or cows fed CSLCFA (starting prepartum, at 1 day in milk (DIM), and 28 DIM). There was a treatment x parity x day inte raction for this contrast (third order polynomial, P < 0.05). The pooled SE of primiparous fed no CSLCFA or CSLCFA was 4.7 and 4.2 pg/ml, respectively. The pooled SE of multiparous cows fed no CSLCFA or CSLCFA wa s 2.5 and 2.4 pg/ml, respectively.

PAGE 113

100 0 5 10 15 20 25 30 35 148121620242832364044 Day postpartumAccumulated progesterone (pg/ml) No CSLCFA CSLCFA A 0 5 10 15 20 25 30 35 40 148121620242832364044 Day postpartumAccumulated progesterone (pg/ml) No CSLCFA CSLCFA B Figure 3.13. Regression plot of accumulated plasma progesterone concentration from d 1 to 46 postpartum of primiparous (A) and multiparous (B) cows fed diets without calcium salts of long chain fa tty acids (CSLCFA) (no CSLCFA) or cows fed CSLCFA (starting prepartum, at 1 day in milk (DIM), and 28 DIM). There was a treatment x parity x day inte raction for this contrast (second order polynomial, P < 0.01). The pooled SE of primiparous cows fed no CSLCFA or CSLCFA was 2.5 and 1.4 pg/ml, re spectively. The pooled SE of multiparous cows fed no CSLCFA or CSLCFA was 2.3 and 1.3 pg/ml, respectively.

PAGE 114

101 Figure 3.14. Regression plot of c oncentration of plasma PGF2 metabolite (PGFM) of primiparous (A) or multiparous (B) cows fed diets without calcium salts of long chain fatty acids (CSLCFA) pr epartum (no CSLCFA plus CSLCFA starting at 1 d in milk (DIM) and 28 DI M), or CSLCFA prepartum. There was a treatment x parity x day interaction fo r this contrast (third order polynomial, P < 0.05). The pooled SE of primiparous cows fed no CSLCFA prepartum or CSLCFA prepartum was 166 and 273 pg/ml, respectively. The pooled SE of multiparous cows fed no CSLCFA prepartum or CSLCFA prepartum was 154 and 271 pg/ml, respectively. 0 1000 2000 3000 4000 1234567891011121314 Day postpartumPGFM (pg/ml) No CSLCFA prepartum CSLCFA prepartumA 0 1000 2000 3000 4000 1234567891011121314 Day postpartumPGFM (pg/ml) No CSLCFA prepartum CSLCFA prepartumB

PAGE 115

102 0 5 10 15 20 25 30 21428Day postpartumTriacylglycerol (% dry liver weight) No CSLCFA CSLCFA prepartum CSLCFA 1 DIM* Figure 3.15. Least squares means fo r hepatic triacylglycerol c oncentration (% of dry liver weight) of cows fed diets without ca lcium salts of long chain fatty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum or at 1 day in milk (DIM). There was a treatment x day interaction (P = 0.04). The asterisk indicates that there was a te ndency for CSLCFA prepartum to differ from no CSLCFA and CSLCFA 1 DIM at 14 DIM (P = 0.084).

PAGE 116

103 A No CSLCFA CSLCFA prepartum CSLCFA 1 DIM__ 7.5 Kb 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 B No CSLCFA ___ CSLCFA prepartum CSLCFA 1 DIM 4.6 Kb 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 C No CSLCFA__ CSLCFA prepartum CSLCFA 1 DIM_ 1.7 Kb 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 D No CSLCFA___ CSLCFA prepartum CSLCFA 1 DIM_ 18S 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 Figure 3.16 Ten micrograms of total cellular RNA isolated from livers of cows fed diets without CSLCFA (lanes 1 to 6), or diets with CSLCFA starting prepartum (lanes 7 to 12) or at 1 DIM (lanes 13 to 18) were subjected to Northern blot analysis. Representitive Northern blot s for IGF-I (A), IGF-II (B), or IGFBP-2 (C) mRNA expression are shown. Within each dietary treatment, two cows are represented at 2 DIM (lanes 1 and 4; 7 and 10; 13 and 16), 14 DIM (lanes 2 and 5; 8 and 11; 14 and 17), and 28 DIM (lanes 3 and 6; 9 and 12; 15 and 18).

PAGE 117

104 Figure 3.17. Least squares means for hepatic IGF-I mRNA abundance of cows fed diets without calcium salts of long chain fa tty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum or at 1 day in milk (DIM). Each bar represents 8 cows per collection period. Values are expressed as ratios over densitometric values for 18S. 0 0.01 0.02 0.03 0.04 0.05 21428Day postpartumIGF-I mRNA A bundance (Arbitrary units / 18S) No CSLCFA CSLCFA prepartum CSLCFA 1 DIM

PAGE 118

105 Figure 3.18. Least squares means for hepatic IGF-II mRNA abundance of cows fed diets without calcium salts of long chain fa tty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum or at 1 day in milk (DIM). ). Each bar represents 8 cows per collection period. Values are expressed as ratios over densitometric values for 18S. 0 0.1 0.2 0.3 0.4 0.5 21428Day postpartumIGF-II mRNA A bundance (Arbitrary units / 18S) No CSLCFA CSLCFA prepartum CSLCFA 1 DIM

PAGE 119

106 Figure 3.19. Least squares means for hepati c IGFBP-2 mRNA abundance of cows fed diets without calcium salts of long ch ain fatty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum or at 1 day in milk (DIM). ). Each bar represents 8 cows per collection pe riod. Values are expressed as ratios over densitometric values for 18S. 0 0.025 0.05 0.075 0.1 0.125 0.15 0.175 0.2 21428Day postpartumIGFBP-2 mRNA Abundance (Arbitrary units / 18S) No CSLCFA CSLCFA prepartum CSLCFA 1 DIM

PAGE 120

107 0 20 40 60 80 13579111315171921232527 Day postpartumHaptoglobin (mg HbB/100 ml) No CSLCFA prepartum CSLCFA prepartum A 0 20 40 60 80 13579111315171921232527 Day postpartumHaptoglobin (mg HbB/100 ml) No CSLCFA prepartum CSLCFA prepartumB Figure 3.20. Regression plot of plasma concentration of haptoglobin of primiparous (A) and multiparous (B) cows fed diets without calcium salts of long chain fatty acids (CSLCFA) prepartum (no CSLCFA plus CSLCFA starting at 1 d in milk (DIM) and 28 DIM) or CSLCFA prepartum. There was a treatment x parity x day interaction for this contra st (third order polynomial, P < 0.05). The pooled standard error (SE) of primiparous cows fed no CSLCFA prepartum or CSLCFA prepartum was 5.5 and 9.0 mg HbB/100 ml (amount of hemoglobin bound by haptoglobin/100 ml of plasma), respectively. The pooled SE of multiparous cows fed no CSLCFA prepartum or CSLCFA prepartum was 5.0 and 9.0 mg HbB/100 ml, respectively.

PAGE 121

108 Figure 3.21. Regression plot of pl asma concentration of ceruloplasmin of cows fed diets without calcium salts of long chain fa tty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum or at 1 day in milk (DIM). This was a third order polynomial (day x day x day, P = 0.003) and the pooled standard error of no CSLCFA, CSLCFA prep artum, and CSLCFA 1 DIM was 7.3, 10.2, and 10.1mg/dl, respectively. 0 5 10 15 20 25 17142128 Day postpartumCeruloplasmin (mg/dl) No CSLCFA CSLCFA prepartum CSLCFA 1 DIM

PAGE 122

109 0 0.1 0.2 0.3 0.4 0.5 0.6 1234 Week postpartumTotal biliruben (mg/dl) No CSLCFA CSLCFA prepartum CSLCFA 1 DIM Figure 3.22. Least squares means for plasma con centration of total biliruben of cows fed diets without calcium salts of long ch ain fatty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum or at 1 day in milk (DIM). Week postpartum was significant (P < 0.001).

PAGE 123

110 0 5 10 15 20 25 30 1234 Week postpartumALT (IU/L) No CSLCFA CSLCFA prepartum CSLCFA 1 DIM Figure 3.23. Least squares means for plasma c oncentration of alanine aminotransferase (ALT) of cows fed diets without calc ium salts of long chain fatty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum or at 1 day in milk (DIM). Week postp artum was significant (P = 0.003).

PAGE 124

111 0 10 20 30 40 50 60 1234 Week postpartumALK (IU/L) No CSLCFA CSLCFA prepartum CSLCFA 1 DIM Figure 3.24. Least squares means for plasma alkaline phosphatase (ALK) concentration of cows fed diets without calcium salts of long chain fatty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum or at 1 day in milk (DIM). Week postpartum wa s significant (P < 0.001).

PAGE 125

112 0 50 100 150 200 1234 Week PostpartumAST (IU/L) No CSLCFA CSLCFA prepartum CSLCFA 1 DIM Figure 3.25. Least squares means for plasma con centration of aspartat e aminotransferase (AST) of cows fed diets without calc ium salts of long chain fatty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum or at 1 day in milk (DIM). Week postp artum was significant (P = 0.01).

PAGE 126

113 0 10 20 30 40 50 1234 Week PostpartumGGT (U/L) No CSLCFA CSLCFA prepartum CSLCFA 1 DIM* Figure 3.26. Least squares means for plasma concentration of gamma glutamyl transferase (GGT) of cows fed diets w ithout calcium salts of long chain fatty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum or at 1 day in milk (DIM). There was a tr eatment x week inte raction (P = 0.01). The asterisk indicates that the treatmen t CSLCFA prepartum is different than no CSLCFA and CSLCFA 1 DIM at wk 4 (P = 0.03).

PAGE 127

114 0 0.5 1 1.5 2 2.5 3 3.5 1234Week postpartumAlbumin (g/dl) No CSLCFA CSLCFA prepartum CSLCFA 1 DIMA 0 0.5 1 1.5 2 2.5 3 3.5 1234Week postpartumAlbumin (g/dl) No CSLCFA CSLCFA prepartum CSLCFA 1 DIMB Figure 3.27. Least squares means for concentratio n of plasma albumin of primiparous (A) or multiparous (B) cows fed diets wit hout calcium salts of long chain fatty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum or at 1 day in milk (DIM). Week postpartum was significant (P = 0.003).

PAGE 128

APPENDIX TESTS OF HETEROGENITY OF REGRESSION

PAGE 129

116Table A-1. Tests of homogeneity of regre ssion and orthogonal contrasts for treatmen t effects on plasma Insulin, IGF-I and accumulated progesterone concentrations of Holstein cows fe d diets without calcium salts of long chain fatty acids (CSLCFA) or diets with CSLCFA starting prepartum, at 1 day in milk (DIM) or at 28 DIM. Error term2 Treatment3 Orthogonal contrasts1 Response Variable Order of regression df MS df MS A B C Insulin 2 1401 0.058 6 0.058 NS4 NS NS IGF-I 2 1404 226 6 1521** ** ** NS Accumulated progesterone, 1 to 77 DIM 3 1448 73.9 9 91.3 NS NS NS Accumulated progesterone, 1 to 46 DIM 2 845 40.5 6 20.6 NS NS NS 1 Orthogonal contrast of curves were the following: A = No CSLCFA vs. CSLCFA, B = CSLCFA prepartum vs. (CSLCFA 1 DIM plus CSLCFA 28 DIM), and C = CSLCFA 1 DIM vs. CSLCFA 28 DIM. 2 Error degrees of freedom (df) and error mean squares (MS) when individual curves were generated for each treatment. 3 Degrees of freedom (df) and mean square (MS) for the difference in residuals betw een fitting one pooled curve vs. fitting an in dividual curve for each treatment. 4 NS = not significant. P < 0.05. ** P < 0.01.

PAGE 130

117Table A-2. Tests of homogeneity of re gression and orthogonal contrasts for treatment effects on plasma PGF2 metabolite (PGFM), haptoglobin, and ceruloplasmin concentrati ons of Holstein cows fed diets without calcium salts of long chain fatty acids (CSLCFA) or diets with CSLCFA starting prepartum, at 1 day in milk (DIM) or at 28 DIM. Error term2 Treatment3 Orthogonal contrasts1 Response Variable Order of regression df MS df MS A B PGFM 3 210 834800 6 224903* NS4 Haptoglobin 3 313 850 6 1488 NS NS Ceruloplasmin 2 314 10.4 4 8.45 NS NS 1 Orthogonal contrast of curves were the following: A = (No CSLCFA plus CSLCFA 1 DIM) vs. CSLCFA prepartum, B =No CSLCFA vs. CSL CFA 1 DIM. 2 Error degrees of freedom (df) and error mean squares (MS) when individual curves were generated for each treatment. 3 Degrees of freedom (df) and mean square (MS) for the difference in residuals betw een fitting one pooled curve vs. fitting an in dividual curve for each treatment. 4 NS = not significant. P < 0.05.

PAGE 131

118Table A-3. Tests of homogeneity of regressi on and orthogonal contrasts for parity e ffects on plasma insulin, IGF-1, accumulated progesterone, PGF2 metabolite (PGFM), haptoglobin, an d ceruloplasmin concentrations of Holstein cows fed diets without calcium salts of long chain fatty acids (CSLCFA) or diets with CSLCFA starting prepartum, at 1 day in milk (DIM) or at 28 DIM. Error term1 Parity2 Response Variable Order of regression df MS df MS Insulin 2 1405 0.058 2 0.215* IGF-I 2 1408 232 2 200 Accumulated progesterone, 1 to 77 DIM 3 1454 74.0 3 136 Accumulated progesterone, 1 to 46 DIM 2 849 40.1 2 165* PGFM 3 213 864500 3 1506833 Haptoglobin 3 316 826 3 4632** Ceruloplasmin 2 316 10.2 2 29.2 1 Error degrees of freedom (df) and error mean squares (MS) when individual curves were generated for each parityt. 2 Degrees of freedom (df) and mean square (MS) for the difference in residuals betw een fitting one pooled curve vs. fitting an in dividual curve for each parity. P < 0.05. ** P < 0.01.

PAGE 132

119Table A-4. Tests of homogeneity of regressi on and orthogonal contrasts for treatment by parity interaction effects on plasma in sulin, IGF-1 and accumulated progesterone concentrations of Holstein cows fed diets without calcium salts of long chain fatty acids (CSLCFA) or diets with CSLCFA starting prep artum, at 1 day in milk (DIM) or at 28 DIM. Error term2 Treatment by parity3 Orthogonal contrasts1 Response Variable Order of regression df MS df MS A B C Insulin 2 1393 0.058 14 0.132** NS4 NS NS IGF-I 2 1396 224 14 934** ** NS Accumulated progesterone, 1 to 77 DIM 3 1436 72.0 21 214** NS ** Accumulated progesterone, 1 to 46 DIM 2 837 39.6 14 85.4** ** NS NS 1 Orthogonal contrast of curves were the following: A = No CSLCFA vs. CSLCFA by parity, B = CSLCFA prepartum vs. (CSLCFA 1 DIM pl us CSLCFA 28 DIM) by parity, and C = CSLCFA 1 DIM vs. CSLCFA 28 DIM by parity. 2 Error degrees of freedom (df) and error mean squares (MS) when individual curves were generated for each treatment and parity. 3 Degrees of freedom (df) and mean square (MS) for the difference in residuals betw een fitting one pooled curve vs. fitting an in dividual curve for each treatment and parity. 4 NS = not significant. P < 0.05. ** P < 0.01.

PAGE 133

120Table A-5. Tests of homogeneity of regressi on and orthogonal contrasts for treatment by parity interaction effects on plasma PG F2 metabolite (PGFM), haptoglobin, and ceruloplasmin concentrations of Holstein cows fed diets without calcium salts of long chain fatty acids (CSLCFA) or diet s with CSLCFA starting prepartum, at 1 day in milk (DIM) or at 28 DIM. Error term2 Treatment by parity3 Orthogonal contrasts1 Response Variable Order of regression df MS df MS A B PGFM 3 201 834079 15 1407712* NS4 Haptoglobin 3 304 830 15 1512* NS Ceruloplasmin 2 308 10.4 10 10.1 NS NS 1 Orthogonal contrast of curves were the following: = (No CSLCFA plus CSLCFA 1 DIM) vs. CSLCFA prepartum by parity, B =No CSLCFA vs. CSLCFA 1 DIM by parity. 2 Error degrees of freedom (df) and error mean squares (MS) when individual curves were generated for each treatment. 3 Degrees of freedom (df) and mean square (MS) for the difference in residuals betw een fitting one pooled curve vs. fitting an in dividual curve for each treatment. 4 NS = not significant. P < 0.05.

PAGE 134

121 LIST OF REFERENCES AbuGhazaleh, A. A., D. J. Schingoethe, A. R. Hippen, and K. F. Kalscheur. 2004. Conjugated linoleic acid increases in milk when cows fed fish meal and extruded soybeans for an extended period of time. J. Dairy Sci. 87:1758-1766. Ackers, M. R. 2002.Page 95 in Lactation a nd the Mammary Gland. Iowa State Press, Ames, IA. Al-Katanani, Y. M., D. W. Webb, and P. J. Hansen. 1999. Factors affecting seasonal variation in 90-day nonreturn rate to first service in lactat ing Holstein cows in a hot climate. J. Dairy Sci 82:2611-2616. Allen, M. S. 2000. Effects of diet on short-te rm regulation of feed intake by lactating dairy cattle. J. Dairy Sci. 83:1598-1624. Armstrong, D. G., T. G. McEvoy, G. Baxter, J. J. Robinson, C. O. Hogg, K. J. Woad, R. Webb, and K. D. Sinclair. 2001. Effect of dietary energy and protein on bovine follicular dynamics and embryo production in vitro: Associations with the ovarian insulin-like growth factor system. Biol. Reprod. 64:1624. Arthington, J. D., S. D. Eicher, W. E. Kunkle, and F. G. Martin. 2003. Effect of transportation and comming ling on the acute-phase prot ein response, growth, and feed intake of newly weaned beef calves. J. Anim. Sci. 81:1120-1125. Atwal, A. S., M. Hidiroglou, J. K. G. Kram er, and M. R. Binns. 1990. Effects of feeding alpha-tocopherol and calcium salts of fa tty acids on vitamin E and fatty acid composition of cow's milk. J. Dairy Sci. 73:2832-2841. Badinga, L., R. J. Collier, W. W. Thatcher, C. J. Wilcox, H. H. Head, and F. W. Bazer. 1991. Ontogeny of hepatic bovine growth hormo ne receptors in cattle. J. Anim. Sci. 69:1925-1934. Bateman, H. G., II, J. N. Spain, and M. R. Ellersieck. 1996. Influence of by-product feeds and tallow on lactation performance of Ho lstein cows during two seasons. J. Dairy Sci. 79:114-120. Bauman, D. E. and J. M. Griinari. 2003. Nutrit ional regulation of m ilk fat synthesis. Ann. Rev. Nutr. 23:203-227. Baumann, H. and J. Gauldie. 1994. The acu te phase response. Immunology Today 15:7480.

PAGE 135

122 Baumgard, L. H., B. A. Corl, D. A. Dwyer, and D. E. Bauman. 2002. Effects of conjugated linoleic acids (CLA) on tissue response to homeostatic signals and plasma variables associated with lipid me tabolism in lactating dairy cows. J. Anim. Sci. 80:1285. Beam, S. and W. Butler. 1997. Energy balance and ovarian follicle development prior to the first ovulation postpartum in dairy cows receiving three levels of dietary fat. Biol. Reprod. 56:133-142. Beam, S. W. and W. R. Butler. 1998. Ener gy balance, metabolic hormones, and early postpartum follicular development in dairy cows fed prilled lipid. J. Dairy Sci. 81:121-131. Beam, S. W. and W. R. Butler. 1999. Effect s of energy balance on follicular development and first ovulation in postpartum dair y cows. J. Reprod. Fertil. 54:411-424. Berman, A., Y. Folman, M. Kaim, M. Mamen, Z. Herz, D. Wolfenson, A. Arieli, and Y. Graber. 1985. Upper critical temperatures and forced ventilation effects for higyielding dairy cows in a sub-tropic al climate. J. Dairy Sci 68:1488-1495. Bilby, T. R., A. Guzeloglu, S. Kamimura, S. M. Pancarci, F. Michel, H. H. Head, and W. W. Thatcher. 2004. Pregnancy and bovine so matotropin in nonlactating dairy cows: I. Ovarian, conceptus, and in sulin-like growth factor syst em responses. J. Dairy Sci. 87:3256-3267. Biochemistry of Lipids, Lipoprot eins, and Membranes. 1996. Page 142 in New Comprehensive Biochemistry. Vol. 31. D. E. Vance and J. E. Va nce, eds, Elsevier, Amsterdam, The Netherlands. Blackshaw, J. K. and A. W. Blackshaw. 1994. Heat stress in cattle and the effect of shade on production and behaviour: A review. Aust. J. of Exp. Agric. 34:285-295. Block, S. S., R. P. Rhoads, D. E. Bauman, R. A. Ehrhardt, M. A. McGuire, B. A. Crooker, J. M. Griinari, T. R. Mackle, W. J. Weber, M. E. Van Amburgh, and Y. R. Boisclair. 2003. Demonstration of a role fo r insulin in the regu lation of leptin in lactating dairy cows. J. Dairy Sci. 86:3508-3515. Brooks, C. G., C. W. Garner, M. E. Gehrke and W. H. Pfander. 1954. The effects of added fat on the digestion of cellulose and protein by ovine rumen microbes. J. Anim. Sci. 13:758-764. Burr, G. O. and M. M. Burr. 1929. A new deficiency disease produced by the rigid exclusion of fat from the diet J. Biol. Chemistry 82:345-367. Burr, G. O. and M. M. Burr. 1930. On the nature and role of the fatty acids essential in nutrition. J. Biol. Chemistry 86:587-621.

PAGE 136

123 Butler, W. R., J. J. Calaman, and B. S.W. 1996. Plasma and milk urea nitrogen in relation to pregnancy rate in lactating da iry cattle. J. Anim. Sci. 74:858-865. Calder, P. C., P. Yaqoob, F. Thies, F. A. Wallace, and E. A. Miles. 2002. Fatty acids and lymphocyte functions. Brit. J. Nutr. 87:S31-S48. Cervantes, A., T. R. Smith, and J. W. Young. 1996. Effects of nicotinamide on milk composition and production in dairy cows fed supplemental fat. J. Dairy Sci. 79:105-113. Chilliard, Y. 1993. Dietary fat and adiopose tissue metabolism in ruminants, pigs, and rodents: A review. J. Dairy Sci. 76:3897-3931. Chilliard, Y., A. Ferlay, R. M. Mansbridge and M. Doreau. 2000. Ruminant milk fat plasticity: Nutritional control of satura ted, polyunsaturated, trans and conjugated fatty acids. Ann. Zootech 49:181-205. Chouinard, P. Y., V. Girard, and G. J. Bri sson. 1997. Lactational re sponse of cows to different concentrations of calcium salts of canola oil fatty acids with or without bicarbonates. J. Dairy Sci. 80:1185-1193. Coulombe, J. J. and L. Favreau. 1963. A new simple method for colorimetric determination of urea. Clin. Chem. 9:102. Delavaud, C., F. Bocquier, Y. Chilliard, D. Keisler, A. Gertler, and G. Kann. 2000. Plasma leptin determination in ruminant s: Effect of nutri tional status and body fatness on plasma leptin concentration assessed by a specific RIA in sheep. J. Endocrinol. 165:519-526. Demetriou, J. A., P. A. Drewes, and J. B. Gin. 1974. Clinical Chemistry. Harper and Row, Hagerstown, MD. Dhiman, T. R., K. V. Zanten, and L. D. Satte r. 1995. Effect of dietary fat source on fatty acid composition of cow's milk. J. Sci. Food Agric. 69:101. Dohmen, M. J. W., J. A. C. M. Lohuis, G. Huszenicza, P. Nagy, and M. Gacs. 1995. The relationship between bacteriological a nd clinical findings in cows with subacute/chronic endometriti s. Theriogenology 43:1379-1388. Douglas, G. N., T. R. Overton, H. G. Bate man, II, and J. K. Drackley. 2004. Peripartal metabolism and production of Holstein co ws fed diets supplemented with fat during the dry period. J. Dairy Sci. 87:4210-4220. Drackley, J. K. 1999. Biology of dairy cows during the transition period: The final frontier. J. Dairy Sci. 82:2259-2273.

PAGE 137

124 Drackley, J. K., D. W. LaCount, J. P. Elliott, T. H. Klusmeyer, T. R. Overton, J. H. Clark, and S. A. Blum. 1998. Supplemental fat and nicotinic acid for Holstein cows during an entire lactation. J. Dairy Sci. 81:201-214. Drackley, J. K., M. J. Richard, D. C. Beitz and J. W. Young. 1992. Metabolic changes in dairy cows with ketonemia in response to feed restriction and dietary 1,3butanediol. J. Dairy Sci. 75:1622-1634. Edmonson, A. J., I. J. Lean, L. D. Weav er, T. Farver, and G. Webster. 1989. A body condition score chart for Holstein dairy cows. J. Dairy Sci 72:68-78. Erickson, P. S., M. R. Murphy, and J. H. Clark. 1992. Supplementation of dairy cow diets with calcium salts of long-chain fatty acids and nicotinic acid in early lactation. J. Dairy Sci. 75:1078-1089. Faldet, M. A. and L. D. Satter. 1991. Feeding heat-treated full fat soybeans to cows in early lactation. J. Dairy Sci. 74:3047-3054. Ferguson, J. D., D. Sklan, W. V. Chalupa, a nd D. S. Kronfeld. 1990. Effects of hard fats on in vitro and in vivo rumen fermenta tion, milk production, and reproduction in dairy cows. J. Dairy Sci. 73:2864-2879. Firkins, J. L. and M. L. Eastridge. 1992. Re placement of forage or concentrate with combinations of soyhulls, sodium bicarbonate or fat for lactating dairy cows. J. Dairy Sci. 75:2752-2761. Foster, L. B. and R. T. Dunn. 1973. Stable reagents for dete rmination of serum triglicerides by a colorime tric hantzsch condensation method. Clin. Chem. 19:338340. Frajblat, M. and W. R. Butler. 2003. Effect of dietary fat prepartum on first ovulation and reproductive performance in lactating dairy cows. J. Dairy Sci. 86 (Suppl. 1):473 (Abstr.). Garcia-Bojalil, C. M., C. R. Staples, C. A. Risco, J. D. Savio, and W. W. Thatcher. 1998. Protein degradability and calcium salts of long-chain fatty acids in the diets of lactating dairy cows: Productive re sponses. J. Dairy Sci. 81:1374-1384. Gillis, M. H., S. K. Duckett, J. R. Sackmann, C. E. Realini, D. H. Keisler, and T. D. Pringle. 2004. Effects of supplemental rume n-protected conjugated linoleic acid or linoleic acid on feedlot performance, carca ss quality, and leptin concentrations in beef cattle. J. Anim. Sci. 82:851-859. Gochman, N. and J. M. Schmitz. 1972. Applica tion of a new peroxide indicator reaction to the specific automated determination of glucose with glucose oxidase. Clin. Chem. 18:943-950.

PAGE 138

125 Grum, D. E., J. K. Drackley, R. S. Younker, D. W. LaCount, and J. J. Veenhuizen. 1996. Nutrition during the dry peri od and hepatic lipid metabolis m of periparturient dairy cows. J. Dairy Sci. 79:1850-1864. Grummer, R. R. 1993. Etiology of lipid-related metabolic di sorders in periparturient dairy cows. J. Dairy Sci. 76:3882-3896. Grummer, R. R. and D. J. Carroll. 1991. Eff ects of dietary fat on metabolic disorders and reproductive performance of dairy cattle. J. Anim. Sci. 69:3838-3852. Hahn, G. L. 1985. Management and housing of fa rm animals in hot environments. Pages 151-174 in Stress physiology in livestock. Vol. 2. M. Yousef, ed. CRC Press, Boca Raton, FL. Harrison, J. H., R. L. Kincaid, J. P. McNamara S. Waltner, K. A. Loney, R. E. Riley, and J. D. Cronrath. 1995. Effect of whole cott onseeds and calcium salts of long-chain fatty acids on performance of lactat ing dairy cows. J. Dairy Sci. 78:181-193. Hawkins, D. E., K. D. Niswender, G. M. Oss, C. L. Moeller, K. G. Odde, H. R. Sawyer, and G. D. Niswender. 1995. An increase in serum lipids increases luteal lipid content and alters the disappearance rate of progesterone in cows. J. Anim. Sci. 73:541-545. Hightshoe, R. B., R. C. Cochran, L. R. Cora h, G. H. Kiracofe, D. L. Harmon, and R. C. Perry. 1991. Effects of calcium soaps of fatty acids on postpartum reproductive function in beef cows. J. Anim. Sci. 69:4097-4103. Hoffman, P. C., R. R. Grummer, R. D. Shav er, G. A. Broderick, and T. R. Drendel. 1991. Feeding supplemental fat and undegraded in take protein to ea rly lactation dairy cows. J. Dairy Sci. 74:3468-3474. Holter, J. E., H. H. Hayes, W. E. Urban, Jr., and A. H. Duthie. 1992. Energy balance and lactation response in Holstein cows supplem ented with cottonseed with or without calcium soap. J. Dairy Sci. 75:1480-1494. Holtman, R. T. 1960. The ratio of trienoic : te traenoic acids in tissu e lipids as a measure of essential fatty acid requirement. J. Nutr. 70:405-410. Jenkins, T. C. and D. L. Palmquist. 1984. Effect of fatty acids or calcium soaps on rumen and total nutrient digestibility of da iry rations. J. Dairy Sci. 67:978-986. Jerred, M. J., D. J. Carroll, D. K. Combs, and R. R. Grummer. 1990. Effects of fat supplementation and immature alfalfa to c oncentrate ratio on la ctation performance of dairy cattle. J. Dairy Sci. 73:2842-2854. Johnson, M. M., and J. P. Peters. 1993. Tech incal note: An improved method to quantify non-esterified fatty acids in bovine plasma. J. Anim. Sci. 71:753-756.

PAGE 139

126 Jorritsma, R., H. Jorritsma, Y. H. Schukke n, and G. H. Wentink. 2000. Relationships between fatty liver and fertility and some periparturient diseases in commercial dutch dairy herds. Theriogenology 54:1065-1074. Kanduce, T. L., A. A. Spector, and R. S. Bar. 1982. Linoleic acid metabolism and prostaglandin production by cultured bovine pulmonary artery endothelial cells. Arteriosclerosis 2:380-389. Khorasani, G. R., P. H. Robinson, G. De Boer, and J. J. Kennelly. 1991. Influence of canola fat on yield, fat percentage, fatty acid profile, and nitrogen fractions in Holstein milk. J. Dairy Sci. 74:1904-1911. Kim, Y. K., D. J. Schingoethe, D. P. Casper, and F. C. Ludens. 1991. Lactational response of dairy cows to increased crude protein with added fat. J. Dairy Sci. 74:3891-3899. Kim, Y. K., D. J. Schingoethe, D. P. Casper, and F. C. Ludens. 1993. Supplemental dietary fat from extruded soybeans and calci um soaps of fatty acids for lactating dairy cows. J. Dairy Sci. 76:197-204. Lambert, M. R., N. L. Jacobson, R. S. Allen, and J. H. Zaletel. 1954. Lipid deficiency in the calf. J. Nutr. 52:259-272. Langhout, D. J., L. J. Spicer, and R. D. Geis ert. 1991. Development of a culture system for bovine granulosa cells: Effects of grow th hormone, estradiol, and gonadotropins on cell proliferation, steroidogenesis, and protein synthesis. J. Anim Sci. 69:33213334. LeBlanc, S. J., T. F. Duffield, K. E. Leslie, K. G. Bateman, G. P. Keefe, J. S. Walton, and W. H. Johnson. 2002. Defining and diagnosi ng postpartum clinical endometritis and its impact on reproductive performan ce in dairy cows. J. Dairy Sci. 85:22232236. Lessard, M., N. Gagnon, D. L. Godson, and H. V. Petit. 2004. Influence of parturition and diets enriched in n-3 or n-6 polyunsat urated fatty acids on immune response of dairy cows during the transition period. J. Dairy Sci. 87:2197-2210. Lewis, G. S. 1997. Uterine health and disorders. J. Dairy Sci. 80:984-994. Liefers, S. C., R. F. Veerkamp, M. F. W. te Pas, C. Delavaud, Y. Chilliard, and T. van der Lende. 2003. Leptin concentrations in re lation to energy balance, milk yield, intake, live weight, and estrus in dairy cows. J. Dairy Sci. 86:799-807. Littell, R. C., G. A. Millike n, W. W. Stroup, and R. D. Wolfinger. 1996.SAS for mixed models. SAS Institute Inc., Cary, NC. Lucy, M. C. 2001. Reproductive loss in high-prod ucing dairy cattle: Where will it end? J. Dairy Sci. 84:1277-1293.

PAGE 140

127 Lucy, M. C., R. L. De La Sota, C. R. Staples, and W. W. Thatcher. 1993. Ovarian follicular populations in lactating dairy cows treated with recombinant bovine somatotropin (sometribove) or saline and fed diets differing in fat content and energy. J. Dairy Sci. 76:1014-1027. Lucy, M. C., J. D. Savio, L. Badinga, R. L. De La Sota, and W. W. Thatcher. 1992. Factors that affect ovarian follicular dynamics in cattle. J. Anim. Sci. 70:36153626. Lucy, M. C., C. R. Staples, F. M. Michel, W. W. Thatcher, and D. J. Bolt. 1991. Effect of feeding calcium soaps to early postpartu m dairy cows on plasma prostaglandin f2alpha, luteinizing hormone, and follicu lar growth. J. Dairy Sci. 74:483-489. Maiga, H. A., D. J. Schingoethe, and F. C. Ludens. 1995. Evaluation of diets containing supplemental fat with different sources of carbohydrates for lacta ting dairy cows. J. Dairy Sci. 78:1122-1130. Makimura, S. C. and N. Suzuki. 1982. Quan titative determination of bovine serum haptoglobin and its elevation in some in flammatory diseases. Jpn. J. Vet. Sci. 44:15-21. Malven, P. V., H. H. Head, R. J. Collie r, and F. C. Buonomo. 1987. Periparturient changes in secretion and mammary uptake of insulin and in concentrations of insulin and insulin-like growth factors in milk of dair y cows. J. Dairy Sci. 70:2254 2265. Markus, S. B., K. M. Wittenberg, J. R. Ingalls, and M. Undi. 1996. Production responses by early lactation cows to whole sunflower seed or tallow supplem entation of a diet based on barley. J. Dairy Sci. 79:1817-1825. Marr, A. L., M. S. Piepenbrink, T. R. Over ton, M. C. Lucy, and W. R. Butler. 2002. The somatotrophic axis and lipid metabolism in transition dairy cows in relation to timing of first postpartum ovulation. J. Dairy Sci 80 (Suppl. 1):263 (Abstr.). Mattos, W. and D. L. Palmquist. 1977. Biohydroge nation and availability of linoleic acid in lactating cows. J. Nutr. 107:1755-1761. The Merck Veterinary Manual. 1997. 8th ed. Merck & Co., Inc., Whitehouse, NJ. Meyer, M. D., P. J. Hansen, W. W. Thatcher, M. Drost, L. Badinga, R. M. Roberts, J. Li, T. L. Ott, and F. W. Bazer. 1995. Extension of corpus luteum lifespan and reduction of uterine secretion of prostagla ndin F2 alpha of cows in response to recombinant interferon-tau. J. Dairy Sci. 78:1921-1931. Moallem, U., Y. Folman, A. Bor, A. Arav, and D. Sklan. 1999. Effect of calcium soaps of fatty acids and administration of soma totropin on milk production, preovulatory follicular development, and plasma and follicular fluid lipid composition in high yielding dairy cows. J. Dairy Sci. 82:2358-2368.

PAGE 141

128 Moallem, U., Y. Folman, and D. Sklan. 2000. Effects of somatotropin and dietary calcium soaps of fatty acids in early lact ation on milk production, dry matter intake, and energy balance of high-yielding dairy cows. J. Dairy Sci. 83:2085-2094. Moallem, U., M. Kaim, Y. Folman, and D. Sklan. 1997. Effect of calcium soaps of fatty acids and administration of somatotropin in early lactati on on productive and reproductive performance of high producing dairy cows. J. Dairy Sci. 80:21272136. National Research Council (NRC). 2001. Nutr ient Requirements of Dairy Cattle. 7th rev. ed. Washington, D. C. Nianogo, A. J., H. E. Amos, M. A. Froets chel, and C. M. Keery. 1991. Dietary fat, protein degradability, and calving season: Effects on nutrient use and performance of early lactation cows. J. Dairy Sci. 74:2243-2255. Oelrichs, W. A., M. C. Lucy, M. S. Kerl ey, and J. N. Spain. 2004. Feeding soybeans and rumen-protected choline to dairy cows dur ing the periparturie nt period and early lactation. 2. Effects on reproduction. J. Dairy Sci. 87 (Suppl. 1):125 (Abstr.). Onetti, S. G., S. M. Reynal, and R. R. Grummer. 2004. Effect of alfalfa forage preservation method and particle length on performance of dairy cows fed corn silage-based diets and tall ow. J. Dairy Sci. 87:652-664. Opsomer, G., Y. T. Grohn, J. Hertl, M. Coryn, H. Deluyker, and A. de Kruif. 2000. Risk factors for post partum ovarian dysfunc tion in high producing dairy cows in Belgium: A field study. Theriogenology 53:841-857. Palmquist, D. L., A. D. Beaulieu, and D. M. Barbano. 1993. Feed and animal factors influencing milk fat compositi on. J. Dairy Sci. 76:1753-1771. Palmquist, D. L. and T. C. Jenkins. 1980. Fat in lactation: Review. J. Dairy Sci. 63:1-14. Palmquist, D. L. and W. P. Weiss. 1994. Bl ood and hydrolyzed feather meals as sources of undegradable protein in high fat diets fo r cows in early lactation. J. Dairy Sci. 77:1630-1643. Pantoja, J., J. L. Firkins, and M. L. Eastri dge. 1996. Fatty acid digestibility and lactation performance by dairy cows fed fats varyi ng in degree of saturation. J. Dairy Sci. 79:429-437. Petit, H. V., R. J. Dewhurst, J. G. Proulx, M. Khalid, W. Haresign, and H. Twagiramungu. 2001. Milk production, m ilk composition, and reproductive function of dairy cows fed different fats. Can. J. Dairy Sci. 81:263-271. Petit, H. V. and H. Twagiramungu. 2002. Reproduction of dairy cows fed flaxseed, Megalac, or micronized soybeans. J. Dairy Sci. 85 (Suppl. 1):312 (Abstr.).

PAGE 142

129 Pickett, M. M., M. S. Piepenbrink, and T. R. Overton. 2003. Effect s of propylene glycol or fat drench on plasma metabolites, liver composition, and production of dairy cows during the periparturient period. J. Dairy Sci. 86:2113-2121. Pires, A. V., M. L. Eastridge, and J. L. Firkins. 1996. Roasted soybeans, blood meal, and tallow as sources of fat and ruminally undegr adable protein in th e diets of lactating cows. J. Dairy Sci. 79:1603-1610. Ried, M. E., J. G. Bieri, P. A. Plack, and E. L. Andrews. 1964. Nutritional studies with the guinea pig: Determination of the li noleic acid requirement. J. Nutr. 82:401-408. Rigout, S., C. Hurtaud, S. Lemosquet, A. B ach, and H. Rulquin. 2003. Lactational effect of propionic acid and duodenal glucose in cows. J. Dairy Sci. 86:243-253. Roberts, A. J., R. A. Nugent, J. Klindt, a nd T. G. Jenkins. 1997. Circulating insulin-like growth factor I, insulin-like growth f actor binding proteins, growth hormone, and resumption of estrus in postpartum cows s ubjected to dietary energy restriction. J. Anim. Sci. 75:1909. Ruppert, L. D., J. K. Drackley, D. R. Bremme r, and J. H. Clark. 2003. Effects of tallow in diets based on corn silage or alfalfa silage on digestion a nd nutrient use by lactating dairy cows. J. Dairy Sci. 86:593-609. Rutter, L. M., R. Snopek, and J. G. Manns 1989. Serum concentrations of IGF-I in postpartum beef cows. J. Anim. Sci. 67:2060 2066. Salado, E. E., G. A. Gagliostro, D. Becu-V illalobos, and I. Lacau-Mengido. 2004. Partial replacement of corn grain by hydrogenate d oil in grazing dairy cows in early lactation. J. Dairy Sci. 87:1265-1278. Salfer, J. A., J. G. Linn, D. E. Otterby, W. P. Hansen, and D. G. Johnson. 1995. Early lactation responses of Holste in cows fed a rumen-inert fat prepartum, postpartum, or both. J. Dairy Sci. 78:368-377. Sanchez, W. K. and E. Block. 2002. Nutriti on and metabolism of fa tty acids in dairy cows. Pages 55-73 in 4th Annual Meeting of Intermountain Nutr. Conf., Utah State Univ., Salt Lake City, UT. Santos, J. E. P., S. O. Juchem, R. L. A. Ce rri, E. J. DePeters, and W. W. Thatcher. 2004. Results of feeding different fatty acids on the cow's transition and reproductive cycle. Page 29-40 in Proc. Southwes t Nutr. & Mgmt. Conf. Tempe, AZ. SAS software statistics. Version 8.2 Edition. 2001. SAS Inst. Inc., Cary, NC. Schingoethe, D. J. and D. P. Casper. 1991. To tal lactational response to added fat during early lactation. J. Dairy Sci. 74:2617-2622.

PAGE 143

130 Schroeder, G. F., J. E. Delahoy, I. Vidaurre ta, F. Bargo, G. A. Gagliostro, and L. D. Muller. 2003. Milk fatty acid composition of cows fed a total mixed ration or pasture plus concentrates replacing corn with fat. J. Dairy Sci. 86:3237-3248. Scott, T. A., R. D. Shaver, L. Zepeda, B. Yandell, and T. R. Smith. 1995. Effects of rumen-inert fat on lactation, reproduction, and health of high producing Holstein herds. J. Dairy Sci. 78:2435-2451. Selberg, K. T., A. C. Dinges, C. R. Staples, and L. Badinga. 2003. Effects of supplemental conjugated linoleic acid a nd trans-octadecenoic fatty acids on the insulin-like growth factor sy stem in periparturient Holstein cows. J. Dairy Sci. 86 (Suppl. 1):139 (Abstr.). Selberg, K. T., A. C. Lowe, C. R. Stap les, N. D. Luchini, and L. Badinga. 2004. Production and metabolic responses of peri parturient Holstein cows to dietary conjugated linoleic acid a nd trans-octadecenoic acids. J. Dairy Sci. 87:158-168. Sewell, R. F. and L. J. McDowell. 1966. Esse ntial fatty acid requirement of young swine. J. Nutr. 89:64-68. Simas, J. M., J. T. Huber, Z. Wu, K. H. Ch en, S. C. Chan, C. B. Theurer, and R. S. Swingle. 1995. Influence of steam-flaked sorghum grain and supplemental fat on performance of dairy cows in early lactation. J. Dairy Sci. 78:1526-1533. Skaar, T. C., R. R. Grummer, M. R. Dentine, and R. H. Stauffacher. 1989. Seasonal effects of prepartum and postpartum fat and niacin feeding on lactation performance and lipid metabolism. J. Dairy Sci. 72:2028-2038. Sklan, D., R. Ashkenazi, A. Braun, A. Devor in, and K. Tabori. 1992. Fatty acids, calcium soaps of fatty acids, and cottonseeds fed to high yielding cows. J. Dairy Sci. 75:2463-2472. Sklan, D., M. Kaim, U. Moallem, and Y. Fo lman. 1994. Effect of dietary calcium soaps on milk yield, body weight, reproductive hormon es, and fertility in first parity and older cows. J. Dairy Sci. 77:1652-1660. Sklan, D., U. Moallem, and Y. Folman. 1991. Ef fect of feeding calcium soaps of fatty acids on production and reproductive responses in high producing la ctating cows. J. Dairy Sci. 74:510-517. Smith, W. A., B. Harris, Jr., H. H. Van Ho rn, and C. J. Wilcox. 1993. Effects of forage type on production of dairy cows suppleme nted with whole cottonseed, tallow, and yeast. J. Dairy Sci. 76:205-215. Son, J., R. J. Grant, and L. L. Larson. 1996. Effects of tallow and escape protein on lactational and reproductive performance of dairy cows. J. Dairy Sci. 79:822-830.

PAGE 144

131 Spicer, L. J., R. K. Vernon, W. E. Tucker R. P. Wettemann, J. F. Hogue, and G. D. Adams. 1993. Effects of inert fat on ener gy balance, plasma concentrations of hormones, and reproduction in dair y cows. J. Dairy Sci. 76:2664-2673. Staples, C. R., J. M. Burke, and W. W. Thatcher. 1998. Influence of supplemental fats on reproductive tissues and performance of l actating cows. J. Dairy Sci. 81:856-871. Staples, C. R., W. W. Thatcher, and J. H. Clark. 1990. Relationship between ovarian activity and energy status during the early postpartum period of high producing dairy cows. J. Dairy Sci. 73:938-947. Stegeman, G. A., D. P. Casper, D. J. Sc hingoethe, and R. J. Baer. 1992. Lactational responses of dairy cows fed unsatur ated dietary fat and receiving bovine somatotropin. J. Dairy Sci. 75:1936-1945. Sutton, J. D., R. Knight, A. B. McAllen, a nd R. H. Smith. 1983. Digestion and synthesis in the rumen of sheep given diets supplemented with free and protected oils. Br. J. Nutr. 49:419-432. Tackett, V. L., J. A. Bertrand, T. C. Je nkins, F. E. Pardue, and L. W. Grimes. 1996. Interaction of dietary fat and acid detergen t fiber diets of lact ating dairy cows. J. Dairy Sci. 79:270-275. Tice, E. M., M. L. Eastridge, and J. L. Fi rkins. 1994. Raw soybeans and roasted soybeans of different particle sizes. 2. Fatty acid utilization by lactating cows. J. Dairy Sci. 77:166-180. Vega, J. R., C. A. Gibson, T. C. Skaar, D. L. Hadsell, and C. R. Baumrucker. 1991. Insulin-like growth factor (IGF)-I and -II and IGF binding proteins in serum and mammary secretions during the dry period and early lactation in dairy cows. J. Anim. Sci. 69:2538-2547. Wang, Y., S. Eleswarapu, W. E. Beal, W. S. Swecker, M. R. Akers, and H. Jiang. 2003. Reduced serum insulin-like growth factor (IG F) I is associated with reduced liver IGF-I mRNA and liver grow th hormone receptor mRNA in food-deprived cattle. J. Nutr. 133:2555-2560. Weiss, W. P. and D. J. Wyatt. 2003. Effect of dietary fat and vitamin E on {alpha}tocopherol in milk from dairy cows. J. Dairy Sci. 86:3582-3591. Wolfenson, D., I. Flamenbaum, and A. Berman. 1988. Hyperthermia and body energy store effects on estrous behavior, concepti on rate, and corpus luteum fuction in dairy cows. J. Dairy Sci 71:3497-3504. Wu, Z., J. T. Huber, S. C. Chan, J. M. Sima s, K. H. Chen, J. G. Varela, F. Santos, C. Fontes, Jr., and P. Yu. 1994. Effect of source and amount of supplemental fat on lactation and digestion in cows. J. Dairy Sci. 77:1644-1651.

PAGE 145

132 Zeron, Y., A. Ocheretny, O. Kedar, A. Borochov, D. Sklan, and A. Arav. 2001. Seasonal changes in bovine fertility: Relation to developmental competence of oocytes, membrane properties and fatty acid composition of follicles. Reproduction 121:447-454.

PAGE 146

133 BIOGRAPHICAL SKETCH Faith Cullens (Fickett) was born in St. Joseph, MI, on July 23, 1980. She attended elementary and high school in South Have n, MI. Faith went on to Michigan State University in 1998, and earned a B.S. degree in zoology. In 2002 she moved to Gainesville, FL, with her future husband, Gus, to begin her Master of Science degree studying dairy nutrition at the University of Florida. After graduation, she will be working as a Dairy Consultant for Carg ill Animal Nutrition in north-west WI.


Permanent Link: http://ufdc.ufl.edu/UFE0010490/00001

Material Information

Title: Effects of the Timing of Initiation of Fat Supplementation on Productive and Reproductive Responses of Periparturient Dairy Cows during Summer
Physical Description: Mixed Material
Copyright Date: 2008

Record Information

Source Institution: University of Florida
Holding Location: University of Florida
Rights Management: All rights reserved by the source institution and holding location.
System ID: UFE0010490:00001

Permanent Link: http://ufdc.ufl.edu/UFE0010490/00001

Material Information

Title: Effects of the Timing of Initiation of Fat Supplementation on Productive and Reproductive Responses of Periparturient Dairy Cows during Summer
Physical Description: Mixed Material
Copyright Date: 2008

Record Information

Source Institution: University of Florida
Holding Location: University of Florida
Rights Management: All rights reserved by the source institution and holding location.
System ID: UFE0010490:00001


This item has the following downloads:


Full Text












EFFECTS OF THE TIMING OF INITIATION OF FAT SUPPLEMENTATION ON
PRODUCTIVE AND REPRODUCTIVE RESPONSES OF PERIPARTURIENT
DAIRY COWS DURING SUMMER















By

FAITH MARIE CULLENS


A THESIS PRESENTED TO THE GRADUATE SCHOOL
OF THE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT
OF THE REQUIREMENTS FOR THE DEGREE OF
MASTER OF SCIENCE

UNIVERSITY OF FLORIDA


2005

































Copyright 2005

by

Faith M. Cullens

































This thesis is dedicated to my wonderful and patient husband, Gus, and to my mom who
has always encouraged me to pursue my dreams.















ACKNOWLEDGMENTS

I would like to thank my committee, Dr. Charles Staples, Dr. Lokenga Badinga,

and Dr. William Thatcher for their time and dedication to my education.

I would like to thank the Davis family for reoccurring scholarship monies, as well

as the dean's office for matching my assistantship. In addition, the IFAS, Graduate

Student Association, and The Animal Sciences Graduate Student Association travel

grants were a great financial assistance in traveling to the American Dairy Science

Association Annual meeting to present my research.

I would also like to thank Catarina Silveria and Myriam Lopez for all of their help

with animal work and with data entry. I would like to thank Todd Bilby, Flavio Silvestre,

Julian Bartolome, Alessandro Sozzi for their help with reproductive management of the

cows. Joyce Hayen played an essential role in pre-trial advising and gave extremely

valuable assistance with lab work. Tremendous help with sample collection and

technical assistance was given by my friends and co-workers including Lucia

Holtshausen, Bruno Amaral, Carlos Alosilla, Marcio Liboni, Tomas Belloso, Sergei

Sennikov, Wimberley Krueger and Elizabeth Johnson. I also would like to thank Dr.

Raymon Littel for his valuable statistical support and Dr. Charles Wilcox for dedicating

many lunch hours to assisting me with statistics. I also would like to thank Dr. John

Arthington for his help analyzing acute phase proteins and Dr. Duane Keisler for

analyzing plasma leptin concentrations. Last but not least, I must thank Mary Russell and

the DRU staff who helped with animal work.
















TABLE OF CONTENTS



A C K N O W L E D G M E N T S ................................................................................................. iv

LIST OF TABLES ....................................................... ............ .............. .. vii

LIST OF FIGURES ......... ......................... ...... ........ ............ ix

ABBREVIATION S ......... .......................... .......... .......... ........... xi

ABSTRACT .............. ..................... .......... .............. xii

CHAPTER

1 INTRODUCTION ............... ................. ........... ................. ... .... 1

2 LITERA TURE REVIEW .......................................................... ..............4

Fatty Acids Defined ................. .. ................ .................. .. .... .... .4
Effects of Supplemental Fat on Feed Intake and Production ......................................5
Tim ing of Initiation of Fat Feeding .................. .............. ............................ ........ 10
Effects of Supplemental Fat on Adipose Tissue Lypolysis...................................12
Effects of Supplemental Fat on Immune Function................. ............................15
Effects of Supplemental Fat on Reproduction................................. ............... 17
A H historical R eview .................. .............................................. 17
The M modern D airy Cow ............................................ .......... ............... 18
Effects of Supplemental Fat on Follicle Development........................................24

3 EFFECTS OF THE TIMING OF INITIATION OF FEEDING CALCIUM
SOAPS OF LONG CHAIN FATTY ACIDS ON PERIPARTURIENT
HOLSTEIN COW S DURING SUM M ER ........................................ .....................45

M materials and M methods ....................................................................... ..................4 5
Cows and Diets ......................... ......... ......... 45
S am ple C collection .............................. .... ...................... .. ........ .... ............4 6
R productive M anagem ent............................................ ........... ............... 47
A nim al H health ........................ .................. ... ............. ......... 49
Sam ple A analysis ........................ ............ ........................ .... 49
Statistical A naly sis ............................................................52




v









R results and D iscu ssion .............................. ......................... ... ........ .... ............54
D ry M atter Intake and D iets ...................................................................... .... 54
Production and B ody W eight ........................................ ......................... 55
Plasm a M etabolites.................. ............................. .... .. .. ... .... ... ..57
Plasm a H orm ones .................. ..................................... .. ........ .... 60
R productive M easurem ents ........................................ .......... ............... 66
H epatic M easurem ents ............................................................. .....................70
Measurements of Immune Status ............................................. .................72
P a rity E ffe cts ................................................................................................. 7 6
C o n c lu sio n ..................................................................................................... 7 7

APPENDIX TESTS OF HETEROGENITY OF REGRESSION................................116

L IST O F R E F E R E N C E S ...................................................................... ..................... 12 1

BIOGRAPH ICAL SKETCH .............................................................. ............... 133















LIST OF TABLES


Table p

2.1 Effects of feeding ruminally inert fat (Ca salts of fatty acids (CSFA) or prilled
fat (PF)) to the same cows throughout the study on DMI, milk production, milk
composition, and body weight (BW) change. ................... ......................... 27

2.2 Effects of feeding oilseeds alone or in combination with other fat sources to the
same cows throughout the experiment on DMI, milk production, milk
composition, and body weight (BW) change. ................... ......................... 33

2.3 Effects of feeding rendered fats to the same cows throughout the study on DMI,
milk production, milk composition and body weight (BW) change......................37

2.4 Effects of feeding ruminally inert fat (Ca salts of fatty acids (CSFA) or prilled
fat (PF)) to the same cows throughout the study on concentration of plasma
horm ones and m etabolites ............................................... ............................. 40

2.5 Effects of feeding oilseeds alone or in combination with other fat sources to the
same cows throughout the study on concentration of plasma hormones and
m etabolites. .......................................... ............................ 43

2.6 Effects of feeding rendered fats to the same cows throughout the study on
concentration of plasma hormones and metabolites. .............................................44

3.1 Ingredient and chemical composition of diets fed to nonlactating cows. ...............78

3.2 Ingredient and chemical composition of diets fed to lactating cows .....................79

3.3 Milk yield, milk composition, postpartum body weight, and postpartum body
condition score. .......................................................................80

3.4 Concentration of plasma hormones and metabolites and day of first ovulation. .....81

3.5 Concentration of plasma PGF2a metabolite (PGFM) the first 14 DIM and the
size and characteristics of the uterus and cervix. ............. ............ ............... 82

3.6 V aginal observations at 21 and 28 D IM ........................................ .....................83

3.7 The number and size of ovarian structures at 21 and 28 DIM ................................84

3.8 Ovarian structures present on the ovaries. ........................................ ....................85









3.9 Concentrations of hepatic triacylglycerol (TAG), and hepatic IGF-I, IGF-II, and
IGF binding protein (BP) -2 mRNA levels ................................... .................86

3.10 Concentration of plasma acute phase proteins and liver enzymes. ..........................87

A-i Tests of homogeneity of regression and orthogonal contrasts for treatment
effects on plasma Insulin, IGF-I and accumulated progesterone concentrations... 116

A-2 Tests of homogeneity of regression and orthogonal contrasts for treatment
effects on plasma PGF2a metabolite (PGFM), haptoglobin, and ceruloplasmin
co n cen tratio n s ............ ............. ....................... ............ ................ 1 17

A-3 Tests of homogeneity of regression and orthogonal contrasts for parity effects on
plasma insulin, IGF-1, accumulated progesterone, PGF2a metabolite (PGFM),
haptoglobin, and ceruloplasmin concentrations. ............................ ... ................ 118

A-4 Tests of homogeneity of regression and orthogonal contrasts for treatment by
parity interaction effects on plasma insulin, IGF-1 and accumulated
progesterone concentrations. .................................................. 119

A-5 Tests of homogeneity of regression and orthogonal contrasts for treatment by
parity interaction effects on plasma PGF2a metabolite (PGFM), haptoglobin,
and ceruloplasm in concentrations. .............................................. ............... 120
















LIST OF FIGURES


Figure pge

3.1 Least squares m eans for m ilk production ............................................................ 88

3.2 Least squares means for body weight. ........................................ ............... 89

3.3 Least squares means for body condition score ....................................................90

3.4 Least squares means for plasma BHBA concentration ........................................91

3.5 Least squares means for plasma NEFA concentration.........................................92

3.6 Least squares means for plasma glucose concentration........................................93

3.7 Least squares means for blood urea nitrogen (BUN) concentration ......................94

3.8 Least squares means for plasma leptin concentration. ...........................................95

3.9 Regression plot of plasma IGF-1 concentration of primiparous (A) and
m ultiparous (B ) cow s .................... .. ...................... .. .. .. .......... .....96

3.10 Regression plot of plasma IGF-1 concentration of primiparous (A) and
m u ltip arou s (B ) cow s ....................................................................... ..................97

3.11 Regression plot of plasma insulin concentration........................ .. ...............98

3.12 Regression plot of accumulated plasma progesterone concentration from d 1 to
77 postpartum ........................................................................99

3.13 Regression plot of accumulated plasma progesterone concentration from d 1 to
46 postpartum .................. ........................................ .. ........ .... 100

3.14 Regression plot of concentration of plasma PGF2, metabolite ...................... 101

3.15 Least squares means for hepatic triacylglycerol concentration.............................. 102

3.16 Ten micrograms of total cellular RNA isolated from livers of cows fed diets
without CSLCFA (lanes 1 to 6), or diets with CSLCFA starting prepartum (lanes
7 to 12) or at 1 DIM (lanes 13 to 18) were subjected to Northern blot analysis.
Representative Northern blots for IGF-I (A), IGF-II (B), or IGFBP-2 (C) mRNA
expression are show n ......................... ...... ................ ............... .... .......... 103









3.17 Least squares means for hepatic IGF-I mRNA abundance ..................................104

3.18 Least squares means for hepatic IGF-II mRNA abundance................................105

3.19 Least squares means for hepatic IGFBP-2 mRNA abundance. ..........................106

3.20 Regression plot of plasma concentration of haptoglobin ................................ 107

3.21 Regression plot of plasma concentration of ceruloplasmin .............. ........... 108

3.22 Least squares means for plasma concentration of total biliruben ..........................109

3.23 Least squares means for plasma concentration of alanine aminotransferase. ........110

3.24 Least squares means for plasma alkaline phosphatase................ ................111

3.25 Least squares means for plasma concentration of aspartate aminotransferase ......112

3.26 Least squares means for plasma concentration of gamma glutamyl transferase.... 113

3.27 Least squares means for concentration of plasma albumin..................................114















ABBREVIATIONS


AI artificial insemination
ALK alkaline phosphatase
ALT alanine aminotransferase
AST aspartate aminotransferase
BW body weight
BCS body condition score
BHBA P-hydroxy butyric acid
BUN blood urea nitrogen
CSFA calcium salts of fatty acids
CSLCFA calcium salts of long chain fatty acids
CL corpus leutem
CLA conjugated linoleic acid
CV coefficient of variation
DM dry matter
DIM days in milk
DMI dry matter intake
EE ether extract
FA fatty acids
FCM fat corrected milk
FSH follicle stimulating hormone
GGT gamma glutamyl transferase
GnRH gonadatropin releasing hormone
LH leutinizing hormone
LA linoleic acid
LNA linolenic acid
NEFA nonesterified fatty acids
NFC non fiber carbohydrates
PGF2a prostaglandin F2,
PGFM 13, 14-dihydro-15-keto-PGF2, metabolite
PHT partially hydrogenated tallow
PUFA polyunsaturated fatty acids
RIA radioimmuno assay
RFM retained fetal membranes
SCC somatic cell count
TAG triacylglycerol
TMR total mixed ration
WCS whole cottonseed















Abstract of Thesis Presented to the Graduate School
of the University of Florida in Partial Fulfillment of the
Requirements for the Degree of Master of Science

EFFECTS OF THE TIMING OF INITIATION OF FAT SUPPLEMENTATION ON
PRODUCTIVE AND REPRODUCTIVE RESPONSES OF PERIPARTURIENT
DAIRY COWS DURING SUMMER


By

Faith M. Cullens

May 2005

Chair: Charles R. Staples
Major Department: Animal Sciences

Primiparous (n = 22) and multiparous (n = 25) Holstein cows were utilized in a

completely randomized block design to determine the effects of timing of the initiation of

feeding Ca salts of long chain fatty acids (CSLCFA) on cow performance the first 100

days in milk (DIM). Four treatments were as follows: control (no CSLCFA) and

CSLCFA supplementation (2% of dietary DM) beginning at 28 d prior to expected

calving date (CSLCFA prepartum), at 1 DIM, or at 28 DIM. Cows fed CSLCFA

beginning in the prepartum period tended (P > 0.05 but < 0.10) to produce more milk

(42.2 vs. 37.1 kg/d) and had (P < 0.05) fewer small (2 to 5 mm) ovarian follicles during

the first estrous synchronization using injection of gonadotropin-releasing hormone (44 +

3 DIM) than cows fed CSLCFA initiated after calving. This milk increase was

accompanied by a lower incidence of disease mastitiss, metritis, or retained fetal

membranes) in the first 10 DIM (8 vs. 43%), elevated concentrations oftriacylglycerol in









the liver dry matter (DM) at 14 + 2 DIM (23.0 vs. 10.4%), lower expression of hepatic

IGF binding protein-2 mRNA (0.08 vs. 0.12 arbitrary units/18S), and elevated

concentrations of plasma bilirubin (0.36 vs. 0.23 mg/dl) compared to cows not fed

CSLCFA prepartum. Expression of hepatic IGF-I and IGF-II mRNA did not differ

among treatment groups. Multiparous cows appeared to benefit more from

supplementation with CSLCFA than primiparous cows in that multiparous cows not fed

CSLCFA at any time during the study had or tended to have lower concentrations of milk

protein, a longer period and greater loss of body weight postpartum, greater

concentrations of plasma beta-hydroxy butyric acid, and lower concentrations of plasma

glucose, leptin, and IGF-I. In addition, multiparous cows fed fat prepartum tended to

have fewer small (2 to 5 mm) and medium (6 to 9 mm) size but more larger size ovarian

follicles (> 10 mm in size) at 21 DIM, more uterine tone at 21 DIM, a smaller cervical os

at 28 DIM, and a slower decrease in plasma concentrations of 13, 14 dihydro-15 keto

prostaglandin F-2a metabolite the first 10 d postpartum than multiparous cows not fed fat

prepartum. Feeding CSLCFA in the prepartum period also benefited primiparous cows

in that they tended to have lower concentrations of plasma fibrinogen. Initiation of fat-

feeding at calving tended to result in fewer and smaller corpus luteum at 21 DIM

compared to those fed CSLCFA prepartum. Estimated day of first ovulation postpartum

based upon two consecutive days of progesterone exceeding 1.0 ng/ml did not differ

among treatment groups (mean of 27 DIM). Multiparous cows fed CSLCFA had greater

concentrations of plasma IGF-I at the time of artificial insemination and conception rate

at first insemination tended to be better for all animals fed CSLCFA (58 vs. 27%)

regardless of day of initiation of CSLCFA supplementation.














CHAPTER 1
INTRODUCTION

The transition from pregnancy to lactation in the dairy cow is a critical phase of the

lactation cycle. The transition period, typically considered the time three weeks

prepartum until three weeks postpartum, is marked by declining dry matter intake (DMI)

as the cow approaches parturition and negative energy status in early lactation. Total

energy intake in early lactation is usually less than what is required for maintenance and

milk production (Staples et al., 1990); therefore the cow must mobilize adipose stores in

the form of nonesterified fatty acids (NEFA) to help support lactation. Nonesterified

fatty acids are removed from circulation by the liver. Once in the liver, NEFA are

utilized for energy; however ruminants have reduced ability to export triglycerides from

the liver and lipids can accumulate causing a metabolic disorder known as fatty liver

(Drackley, 1999).

Supplemental fats can increase the energy density of the diet and reduce an energy

deficit in early lactation. However milk production often increases when fats are added

to the diet, resulting in no improvement of energy status. In addition, fat feeding can

often result in a depression of DMI (Allen, 2000). The mechanisms by which

supplemental fat depresses DMI are not apparent but could involve negative effects on

ruminal fermentation and gut motility, release of gut hormones, oxidation of fat in the

liver, and palatability of diets containing added fat (Allen, 2000). Feeding supplemental

fat in the form of Ca salts of fatty acids (CSFA) makes the fat partially inert in the rumen

and sometimes can prevent a depression in DMI. Feeding CSFA can allow specific fatty









acids (FA) to escape ruminal biohydrogenation which are then available in the small

intestine for absorption and utilization. The modern dairy cow may be deficient in the

essential FA, linoleic acid (LA) (Sanchez and Block, 2002). Providing LA in the form of

CSFA may reduce a deficiency and act on target tissues in addition to increasing the

energy density of the diet.

Supplemental fats have proven beneficial to reproductive efficiency of lactating

dairy cows as well (Staples et al., 1998). Although reproductive performance is strongly

associated with energy status (Staples et al., 1990), dietary fats can provide FA precursors

for cholesterol and prostaglandin production, which have an affect on ovarian function,

uterine function, and conception rates. Immune reactions have also been shown to be

modulated by the diet including the FA composition by influencing cellular

communication and activation through the synthesis of prostaglandins (Calder et al.,

2002). Decreased incidence of disease in early lactation can result in increased milk

production throughout the lactation and an increase in reproductive efficiency. The

amount of a particular FA (e.g., LA) stored in the target tissue may control prostaglandin

synthesis.

The modern dairy cow in the United States is most productive between the

temperatures of 5 and 150C and will begin to experience slight losses in milk production

between 15 and 250C (Hahn, 1985). Above 260C, dramatic losses in production can

occur although the humidity index will alter this upper critical temperature (Berman et

al., 1985). In an attempt to dissipate additional body heat, the cow will increase her

respiration rate, increase sweating rate, increase blood flow to skin, and decrease energy

intake (Blackshaw and Blackshaw, 1994). Because fat has a low heat increment









associated with feeding, has a high energy density, and is utilized with high efficiency, it

is an ideal feed additive during periods of heat stress. Reducing heat stress in lactating

dairy cattle can increase milk production and reproductive efficiency. Heat stress can

alter hormonal profiles and reduce the duration and intensity of estrus (Wolfenson et al.,

1988). Using a large group of Isreali cows, Zeron and coworkers (2001) reported

lowered conception rates in summer versus winter months which may have been due to

differences in follicular dynamics and changes in the biological membranes. In the

winter, more follicles per ovary and embryo development to the blastocyst stage, the two-

to four-cell stage, and the morula stage were improved compared to summer months. In

addition, the proportion of FA in the phospholipids of oocytes, granulosa cells, and

follicular fluid was increased in the summer whereas the proportion of polyunsaturated

FA in the phospholipids of oocytes and granulosa cells was increased in the winter. A

study examining pregnancy rates in Florida and Georgia herds using DHIA records

reported that high milk production exacerbated the already great drop in non-return rates

during the summer months (Al-Katanani et al., 1999).

The objective of this thesis was to evaluate the timing of initiation of Ca salts of

long chain FA on the production, plasma hormones and metabolites, and reproduction of

periparturient Holstein cows in summer.














CHAPTER 2
LITERATURE REVIEW

Fatty Acids Defined

Linoleic acid (LA), the major fatty acid (FA) in most oilseeds, and linolenic acid

(LNA), the major FA in fresh forages, are considered essential FA because they cannot

be synthesized by mammals or ruminal microorganisms. They lack the A 12 and A 15

desaturase enzymes to convert oleic acid to LA and LNA. Both LA and LNA are

polyunsaturated FA (PUFA) in that they have more than one point of unsaturation, or

double bond. The shorthand notation system for identifying FA includes a description of

the number of carbons present within the chain, the number of double bonds, and the

location and configuration of the double bonds. In addition, the co-numbering system

begins numbering carbons starting at the methyl end of the FA. For example, LA is

notated as C18:2, co-6 because it has 18 carbons, two double bonds, and the first double

bond is at the sixth carbon from the methyl end. The LNA is designated C18:3, co-3.

Lastly, FA can be classified in the co-9 category such as oleic acid, C18:1. Omega

families cannot be interconverted.

Fat supplementation is commonly used to increase the energy density of the diet of

lactating dairy cows. During the first few weeks of lactation, dairy cows are limited by

nutrient intake by which to meet the demands of lactation. Early postpartum dry matter

intake (DMI) is limited by ruminal fill and does not peak until 10 to 14 wk postpartum,

while milk production usually peaks at 4 to 8 wk postpartum (National Research Council

[NRC], 2001).









Feeding diets with 3 to 5% added fat can increase energy intake without having a

major negative effect on fiber digestion or milk fat (Palmquist and Jenkins, 1980).

Supplying energy in the form of fat in place of carbohydrate can reduce microbial protein

production since carbohydrates are the primary energy source for ruminal microbes. On

the other hand, increasing the energy concentration of the diet by increasing the starch to

an excessive concentration can have detrimental effects on digestion and animal health.

Feeding excessive unsaturated FA, which have a toxic effect on ruminal microbes,

may lead to depressed fiber digestion and milk fat production (Brooks et al., 1954;

Jenkins and Palmquist, 1984). Manufacturing Ca salts of fatty acids (CSFA) can make

them partially inert in the rumen and can increase the energy density of the diet without

hindering forage digestion (Jenkins and Palmquist, 1984). Preformed soaps of FA should

not dissociate in the rumen, but dissociate in the abomasum at a low pH. The FA are then

available in the small intestine for absorption and utilization by the body (Jenkins and

Palmquist, 1984).

The studies reviewed in this thesis have been limited to those in which cows were

fed the same experimental diets for the duration of the study. Important aspects of each

study are summarized in Table 2.1 (ruminally inert fats), Table 2.2 oilseedss) and Table

2.3 (rendered fats).

Effects of Supplemental Fat on Feed Intake and Production

Dry matter intake is often affected by the addition of fat to the diet (Allen, 2000;

Jerred et al., 1990) The type of fat fed as well as the type and amount of forage will have

an effect on the extent to which DMI is affected (Allen, 2000). Although the

mechanisms are unclear, intake may be depressed when supplemental fats are fed due to

decreased palatability, ruminal fill due to inhibition of fiber digestion, the metabolic









regulation of the gut hormone cholecystokinin on the brain satiety centers, and an

increased rate of FA oxidation in the liver that can alter signals generated by hepatic

vagal afferent nerves to brain centers signaling satiety (Allen, 2000). In addition, PUFA

that reach the small intestine may decrease gut motility which could decrease DMI

(Drackley et al., 1992). In a review of several studies, Allen (2000) reported that there

was a linear decline in DMI such that for every 1% inclusion of tallow or CSFA in the

diet there was a reduction of DMI of 1.2 and 2.5% respectively. Allen also reported that

oilseeds negatively affected intake, although in a quadratic fashion with 2% of dietary FA

coming from oilseeds resulting in maximum intake suppression.

In this review of studies where CSFA were fed to the same animals throughout the

study (Table 2.1), the majority of experiments report no effect on DMI when CSFA were

fed at 1.8 to 5% of dietary dry matter (DM) (Atwal et al., 1990; Chouinard et al., 1997;

Erickson et al., 1992; Firkins and Eastridge, 1992; Holter et al., 1992; Moallem et al.,

2000; Palmquist and Weiss, 1994; Schroeder et al., 2003; Skaar et al., 1989; Spicer et al.,

1993). However, several studies reported a depression in DMI when cows were fed

CSFA in diets in which alfalfa (silage or hay) was the sole forage source (Chouinard et

al., 1997; Harrison et al., 1995; Jerred et al., 1990; Simas et al., 1995). Interestingly, a

few studies reported DMI was depressed early in the experiment; however the effect

disappeared after the cows consumed the diets for a longer period of time (Beam and

Butler, 1998; Chouinard et al., 1997; Garcia-Bojalil et al., 1998), suggesting that there

may be an adaptation period.

Depression of DMI was less commonly reported when cows were fed oilseeds as a

fat source throughout the duration of the study. In two studies by Harrison and









coworkers (1995), intake was depressed when early lactation cows consumed diets of

12% whole cottonseed (WCS) and 2.7 or 5% of DM as CSFA (Table 2.2). In one of

those studies, DMI was depressed when a diet of 12% WCS without CSFA was fed.

When high amounts of whole soybeans (18% of dietary DM and 6.2% dietary ether

extract (EE)) were fed in a diet containing 30% corn silage and 20% alfalfa silage, DMI

was depressed 2.3 kg/d (9.7% of DMI) in comparison to controls (3.2% dietary EE).

However the feeding of soybeans in the ground roasted form (18% of dietary DM) did

not affect DMI (Pires et al., 1996). The roasting process may reduce exposure of the oil

to the rumen microbes and therefore reduce the negative influence on rumen function.

Escape of LA to the small intestine was increased in cows fed roasted versus raw

soybeans (Dhiman et al., 1995; Tice et al., 1994). Similarly, the addition of rolled

safflower seeds at 10% of dietary DM in combination with bST injections (5.5% dietary

EE) depressed DMI by 4.6 kg/d (19.2% of DMI) in comparison to control cows receiving

bST injections (2.3% dietary EE) in a diet of 25% corn silage and 25% alfalfa hay (DM

basis). However, the replacement of safflower seeds with rolled sunflower seeds (6.2%

dietary EE) in combination with bST injections did not affect DMI (Stegeman et al.,

1992).

In this review of studies where rendered fats were fed to the same animals

throughout the experiment, only two studies reported a suppression in DMI in

comparison to the control or similar treatment without fat (Table 2.3). Bateman and

coworkers (1996) reported a depression of 2.6 kg/d (10% of DMI) when tallow (2% of

dietary DM) was fed in a 33% NDF diet in the winter (3.5 and 5.4% dietary EE for fat-

supplemented and control diets, respectively). However, tallow feeding had no effect on









DMI when included in the summer or in a diet with 40% NDF. Lastly, when lactating

cows were fed diets of 0 or 3% tallow (DM basis) in a ration comprised of 33% alfalfa

haylage and 17% corn silage, DMI was depressed 0.9 kg/d (3.6% of DMI) and 1.3 kg/d

(5.4% of DMI) with the addition of 0 or 5% escape protein supplement, respectively (Son

et al., 1996).

Changes in milk production can accompany changes in DMI. Often, milk

production is increased due to increased energy intake when supplemental fats are fed,

particularly in early lactation when the dairy cow must depend on her body reserves to

help satisfy energy requirements for maintenance and lactation. Of 28 studies in which

ruminal inert fat was fed to cows in continuously applied treatments, eleven studies did

not report an increase in production of milk or fat-corrected milk (FCM) (Table 2.1).

Reasons for the discrepancies between studies may be due in part to the day postpartum

that fat feeding was initiated, the duration of the study, feed quality, amount of milk

production, and combinations of these factors. In one study that did not report an

increase in milk production, cows were supplemented CSFA while on pasture (Schroeder

et al., 2003). In another study that did not report an increase in milk production, a 0.45

kg drench of CSFA was administered for only 4 d postpartum (Pickett et al., 2003).

Cervantes and coworkers (1996) did not report an increase in milk production when

feeding 0.4 kg/d of CSFA but the cows were in midlactation (average of 112 DIM) and

were only fed diets for 38 d. Additionally, two studies that did not report an increase in

milk production were already feeding high concentrate diets (> 61% of dietary DM) that

included > 8.5% WCS and the additional energy provided by dietary fat may not have

been beneficial (Simas et al., 1995; Spicer et al., 1993).









Similarly, in 11 of 17 studies in which oilseeds were fed to the same animals

throughout the experiment, milk production or FCM increased (Table 2.2). Two studies

that did not report an increase in milk production were already feeding high concentrate

diets (> 60% of dietary DM) (Khorasani et al., 1991; Markus et al., 1996). Another study

reported DMI depression of 9.7% when whole soybeans were fed, which may have

contributed to the unaffected milk production (Pires et al., 1996).

When rendered fats were fed in continuous studies, six of 13 studies reported an

increase in milk production or FCM yield (Table 2.3), and interestingly there was no

suppression in DMI. Reasons for this variation in response across studies were not

apparent.

Milk protein concentration was commonly reported to decline when ruminally inert

fats or oilseed were fed (Tables 2.1 and 2.2) but less often when rendered fats were fed

(Table 2.3). Although the mechanism is unclear, it could be due, in part, to less

glucogenic precursors being consumed as starch is replaced with lipid. Less glucogenic

precursors present in the diet is associated with decreased milk protein concentration

(Rigout et al., 2003).

The response of milk fat concentration to supplemental fat seems to be dependent

upon many factors. About 50% of the fat found in milk is synthesized in the mammary

gland from acetate and butyrate, while the other 50% comes directly from fat absorbed

from the blood (Ackers, 2002). Palmquist and coworkers (1993) published an equation

predicting an increase in milk fat concentration of 0.18% from feeding an additional 0.5

kg of fat daily. Conversely, milk fat depression is often seen when fat is fed in diets in

which corn silage was the sole forage source, resulting in a more acidic ruminal









environment than when fed in combination with alfalfa hay or haylage which has a

greater buffering capacity (Onetti et al., 2004; Ruppert et al., 2003; Smith et al., 1993).

This is thought to occur when PUFA are biohydrogenated in the rumen under acidic

conditions to trans C18:1 FA rather than to C18:0. The enzymes responsible in the

mammary gland to synthesize the short and medium chain fatty acids found in milk may

be inhibited by these trans C18:1 FA, especially trans-10 C18:1, resulting in decreased

milk fat concentration (Bauman and Griinari, 2003). In this review of studies in which

supplemental fat was fed to the same cows throughout the experiment, an increase in

milk fat concentration was more consistent when cows were fed CSFA in comparison to

oilseeds or rendered fats.

Accompanying an increase in milk production due to fat supplementation may be

an increase in 4% FCM yield (Erickson et al., 1992; Moallem et al., 2000; Salado et al.,

2004). However, if milk fat concentration is unchanged or is depressed by fat

supplementation, yield of 4% FCM may not significantly increase despite an increase in

milk production (AbuGhazaleh et al., 2004; Pantoja et al., 1996; Selberg et al., 2004).

Timing of Initiation of Fat Feeding

Few studies in the literature examine when to start feeding fat during the

periparturient period. If fat supplementation is begun in the dry period, benefits can be

expected due to the adjustment of the animal's palate and ruminal microflora before the

onset of the lactation. Skaar and workers (1989) fed 40 multiparous cows diets of 0 or

5% prilled fat containing 50% forage (equal amounts of corn silage and alfalfa silage,

DM basis) beginning at 17 d before expected calving date and continued through 105 d in

milk (DIM). Intake did not differ between fat-fed and control cows yet overall milk

production tended to increase for cows fed fat. During the cool season, milk production









did not differ between treatments, however, during the warm season milk yield increased

from 34.5 to 44.3 kg/d. Although cow performance was not negatively affected, liver

biopsies revealed that cows fed fat beginning prepartum tended to have increased total

hepatic lipid concentrations in comparison to controls (27.5 vs. 26.1%, DM basis at 1

DIM and 29 vs. 24%, DM basis at 5 weeks postpartum). Plasma nonesterified fatty acid

(NEFA) concentrations did not differ between treatments indicating that the additional

lipid in the liver may have been from dietary origin.

Waiting until calving to start feeding fat is another approach often tried because of

the extra costs associated with fat supplementation. University of Florida workers

(Garcia-Bojalil, et al., 1998) fed multiparous cows corn silage/alfalfa hay-based diets

containing 0 or 2.2% calcium salts of palm oil beginning at calving and continuing

through 120 DIM. Fat-fed cows began to produce more milk (-2 kg/d) after 3 weeks of

supplementation and continued to produce more milk throughout the experiment.

Allowing cows to get through the period of negative energy balance before adding

fat to the diet is one more strategy employed. Schingoethe and Casper (1991)

summarized five studies in which whole sunflower or extruded soybeans were fed

beginning at 4 weeks and continued through 16 weeks of lactation. Cows produced an

average of 0.9 kg/d more milk while consuming the seeds, however, there was still a 3 to

4 week delay before a milk response was evident.

Salfer and coworkers in Minnesota (1995) evaluated when to initiate the feeding of

partially hydrogenated tallow (PHT). Sixty-three animals were assigned to treatment at

14 d before expected calving date and remained on the same treatment until 151 DIM.

Four dietary treatments were the following: 1) no PHT prepartum or postpartum, 2) 1%









PHT prepartum and 2% PHT postpartum, 3) 0% PHT prepartum and 2% PHT

postpartum, and 4) 0% PHT prepartum and 2% PHT beginning at 35 DIM. Dry matter

intake and milk production did not differ among treatments. When evaluating the first 35

d of lactation alone, cows fed fat beginning at calving had a 3.5 kg/d advantage of 3.5%

FCM yield in comparison to cows fed fat beginning in the prepartum period, due in part

to differences in the concentration of milk fat between the two treatments. Plasma NEFA

concentrations, days to first estrus, pregnancy rate, days open, and incidence of disease

did not differ among treatments.

Effects of Supplemental Fat on Adipose Tissue Lypolysis

When the energy needed for maintenance and lactation is greater than the energy

provided in the diet, the dairy cow will begin to mobilize her body fat stores to lessen the

energy deficit. Nonesterified fatty acids are released into the blood by adipose tissue and

transported to hepatic and non-hepatic tissues. Once in the liver, NEFA have the three

following fates: 1) oxidized to carbon dioxide to provide energy, 2) partially oxidized for

energy but also producing ketone bodies such as P-hydroxy butyric acid (BHBA) that

serve as fuel for other tissues, or 3) reconverted to triglycerides and stored (Drackley,

1999). When fat is added to the diet, plasma concentrations of NEFA routinely increase

(Chilliard, 1993; Drackley, 1999; Grummer and Carroll, 1991). In a review of 50

treatment comparisons, Chilliard (1993) reported an average increase in concentration of

plasma NEFA of 41 tM (P < 0.005) over controls when supplemental fat was fed.

Likewise, Drackley (1999) reported an average increase in concentration of plasma

NEFA of 81 tM over controls when supplemental fat was fed in reviewing seven studies.

This increase due to dietary fat is much less than what is typically observed during the









transition period when NEFA concentrations may increase up to 1 mM or more

(Grummer, 1993).

When NEFA concentrations in blood are high, as during the periparturient period,

removal of NEFA by the liver may exceed the oxidative capacity of the liver and, when

in combination with the low rate of export of triacylglycerol (TAG) out of the liver in

ruminants, TAG accumulates in the liver (Grummer, 1993). Mechanisms to decrease

hepatic lipid accumulation in cows fed fat might include decreased hepatic uptake of

NEFA, increased oxidation of NEFA in the liver, decreased hepatic esterification of

NEFA, or increased export of TAG from the liver (Grum et al., 1996).

Illinois workers (Grum et al., 1996) conducted a study in which cows were fed a

control diet (80% oat hay, DM basis), a high concentrate diet (51% oat hay, DM basis),

or a high fat diet (6.5% of DM as EE) for 50 d prepartum and a common diet postpartum.

Cows fed fat during the prepartum period tended to have decreased plasma NEFA

concentrations early postpartum and had decreased liver TAG concentrations at d 1 of

lactation. They also reported a positive correlation between concentrations of plasma

NEFA at 3 d prepartum and the concentration of TAG in the liver at 1 d postpartum.

However, a similar study at Illinois (Douglas et al., 2004) in which cows were fed a

moderate non-fiber carbohydrate (NFC) control diet, a low NFC diet with 4% choice

white grease (Qual-Fat) prepartum (DM basis), and a moderate NFC diet with 4%

choice white grease (DM basis) beginning at 60 d prepartum revealed no treatment

differences in DMI, milk production, plasma concentration of NEFA, or total hepatic

lipid or TAG at 1 DIM.









During the periparturient period, the adipose tissue transfers from an anabolic state

to a catabolic state. Tissue mobilization increases because the nutrient demands of the

fetus and placenta are high and DMI declines during the last weeks of pregnancy

(Grummer, 1993). Endocrine secretions have a major impact on lipogenesis and

lipolysis. Insulin increases glucose uptake by the cell membrane and increases lipogenic

enzymes to stimulate FA and TAG synthesis. Staples et al. (1998) reviewed 17 studies

that reported insulin concentrations from fat-supplemented cows and found mixed results.

In 8 studies, concentration of plasma insulin of cows fed fat were significantly depressed

in comparison to controls, however differences were eliminated when adjusted for energy

status of the animals.

Leptin is a hormone synthesized by adipose tissue that is positively regulated by

adiposity and negatively by undernutrition. High concentrations of plasma leptin are

associated with decreased feed intake and increased energy expenditure. Low

concentrations of plasma leptin are associated with increased appetite and energy

conservation. Concentrations of leptin are decreased around the time of parturition in

concurrence with negative energy balance and a reduction in adipose stores and may be

mediated by the reduction in plasma insulin (Block et al., 2003). Circulating

concentrations of plasma leptin were not different between beef heifers fed a basal diet or

diets with supplemental fat (4% corn oil or 2% Ca salts of conjugated LA (CLA), DM

basis) for 32 or 60 d before slaughter (Gillis et al., 2004). However, leptin concentrations

in adipose tissue were greater for heifers supplemented with 4% corn oil versus heifers

fed the basal diet or 2% Ca salts of CLA (0.28, 0.18, and 0.17 pg/pg protein,

respectively). Likewise, concentration of plasma leptin of late lactation dairy cattle









abomasally infused with cis-9, trans-11 or trans-10, cis-12 CLA isomers were not

different (Baumgard et al., 2002).

Effects of Supplemental Fat on Immune Function

In response to an activated immune system, the liver will produce acute phase

proteins such as ceruloplasmin, fibrinogen, and haptoglobin (Baumann and Gauldie,

1994). Haptoglobin is responsible for preventing the loss of body iron and concentrations

are normally undetectable in bovine blood unless there is tissue damage. Ceruloplasmin

is involved with copper transport and concentrations will increase due to an inflammatory

response of the cow. Fibrinogen is involved with blood clotting and the formation of the

fibrin matrix for tissue repair. Increased fibrinogen concentrations are detected during

internal hemorrhage or tissue damage. Arthington and coworkers (2003) looked at the

response of newly weaned beef calves to the stresses of transportation and co-mingling

and reported increased concentrations of plasma fibrinogen, ceruloplasmin, and

haptoglobin.

Immune reactions have been shown to be modulated by the diet, including the

PUFA composition of the diet (Calder et al., 2002). Mechanisms involved in regulation

are not yet understood, but evidence exists that PUFA composition of the diet influences

cellular communication and activation through the synthesis of prostaglandins, tumor

necrosis factor-a, and interferon-y (Calder et al., 2002). Linoleic acid can be converted to

arachidonic acid, the precursor for prostaglandin E2 and leukotriene B4 which are pro-

inflammatory mediators. Similarly, LNA can be converted to eicosapentaenoic acid, the

precursor for the synthesis of the inflammatory mediators prostaglandin E3 and

leukotriene B5. Lessard et al. (2004) evaluated cellular immune functions of dairy cows

fed supplemental fat during the transition period. Cows were fed diets of 2.7% Ca salts









of palm oil (Megalac), 5.9% flaxseed (n-3 FA) or 9.4% micronized soybeans (n-6 FA)

from 6 wk prepartum to calving. From calving to 6 wk postpartum, cows were fed diets

of 4.7% Megalac, 9.7% flaxseed, or 20.3% micronized soybeans. Serum antibody

response to ovalbumin injections during the prepartum period did not differ among

treatments. The lymphocyte response of blood mononuclear cells to mitogenic

stimulation was lower in cows fed soybeans than in those receiving flaxseed or Megalac.

The authors concluded that cellular immune functions were modulated around

parturition; however feeding diets rich in n-3 or n-6 FA did not have a major impact on

cellular immune function.

Immediately postpartum up to 90% of cows develop mild endometritis (Lewis,

1997). It is suggested by many researchers that a compromised immune system

involving reduced effectiveness of neutrophilic movement and pathogen destruction may

be the reason that some cows spontaneously recover while others develop severe uterine

infections that reduce fertility (Lewis, 1997). Evaluation of cervical discharge using

vaginoscopy can be used as a diagnosis tool of uterine bacterial infection (Dohmen et al.,

1995). Clinical endometritis as described by a purulent or foul discharge after 20 d

postpartum or a mucopurulent discharge after 26 d postpartum was associated with a

reduction of pregnancy rates (LeBlanc et al., 2002). Abnormal vaginal discharge has

been correlated with a delay in the first postpartum ovulation (Opsomer et al., 2000).

Furthermore, if first ovulation occurs in the presence of a uterus with heavy

contamination, it can lead to prolonged luteal phases which is also associated with lower

fertility (Opsomer et al., 2000).









Effects of Supplemental Fat on Reproduction

A Historical Review

It is well established in nonruminants that animals with essential FA deficiencies

have poor skin and hair, low growth rates, and reduced reproductive performance. In

early studies, the essentiality of LA was documented primarily in nonruminants by

causing then curing symptoms of deficiency. In the late 1920's Burr and Burr (1929)

established that dietary fat was essential to the growing rat. After 70 d on purified diets,

rats without lipid in the diet experienced dandruff, hair loss, cessation of growth,

abnormal kidneys, blood in the urine, prolapsed penis, and irregular ovulation. All

animals died after 120 to 230 d without dietary fat. The following year, Burr and Burr

(1930) sought to identify specific FA responsible for normal reproductive function in

rats. Growing rats were fed lipid-free diets and upon weight loss, individual lipid sources

were supplemented at 1% of the diet. Of 22 females fed fat-free diets, 13 were not

cycling or cycling irregularly. When three ovulatory rats that had been fed fat-free diets

were bred, two produced litters; however no young lived more than a few hours. When

four of the nonovulatory females were given five drops of either corn (41% LA), olive

(7% LA), linseed (59% LA), or coconut (1% LA) oil daily, all resumed ovulation except

for the rat supplemented with coconut oil. Six of the irregularly ovulating females fed

fat-free diets were fed two drops of cod liver oil daily. After 4 wk all rats were cycling

normally, were bred and produced normal litters. In response to their results, the authors

stated, "...the resumption of ovulation is so rapid that growth has hardly begun.

Synthesis of ovarian hormone ceases when fatty acids are eliminated from the diet." It

was many years later that FA deficiency was studied in farm animals. In 1954 it was

determined that the preweaned calf also requires FA (Lambert et al., 1954). Preruminant









calves were fed isocaloric, purified diets of synthetic milk devoid of lipid with or without

hydrogenated soybean oil and lecithin. Calves fed diets without lipid developed

symptoms of deficiency including scaly dandruff, long dry hair, hair loss, diarrhea, and

low weight gain. Additionally, guinea pigs fed a basal fat-free diet for eight months had

an average weight of 254 28 g, incidence of dermatitis was 75% and mortality rate was

25% while guinea pigs raised on the same diet and supplemented with 0.4% methyl

linoleate (1.31% of calories) had an average weight of 382 15 g and had no dermatitis

or mortality (Ried et al., 1964). Lastly, growing male pigs were fed 0, 0.25, 0.5, 1.0, 2.0,

and 4.0% of dietary calories as LA using corn oil for ten wk (Sewell and McDowell,

1966). Concentration of LA in scrotal fat reflected dietary intake after five and ten wk on

diet. Skin lesions appeared on pigs receiving 0, 0.25, and 0.5% of dietary calories as LA

and they were then supplemented with methyl linoleate or methyl oleate at 1.0% of

dietary calories. Skin lesion disappeared in pigs supplemented with methyl linoleate and

remained on those supplemented with methyl oleate.

It has been documented in nonruminants, including the rat (Holtman, 1960), the

guinea pig (Ried et al., 1964), and the pig (Sewell and McDowell, 1966) that a ratio of

C20:3 to C20:4 in tissues/serum that exceeds 0.4 is indicative of a LA deficiency. The

rational behind this ratio as an indicator of LA deficiency is that the synthesis of C20:3 n-

9 from oleic acid increases when LA is deficient because of enzyme competition.

The Modern Dairy Cow

Quantification of LA available for use by the adult ruminant is difficult to predict

because of ruminal biohydrogenation. The extent of ruminal biohydrogenation is

variable and dependent upon many factors including the diet and ruminal conditions,

however it is estimated by Chilliard et al. (2000) that 80% of dietary LA is









biohydrogenated. Feeding ruminally inert fats, such as CSFA, will partially protect FA

from biohydrogenation, allowing greater escape to the small intestine.

Using the fat sub-model of the Cornell-Penn-Miner (CPM)-Dairy model, Sanchez

and Block (2002) suggested that the amount of LA excreted in 45.5 kg of milk daily

exceeds the post ruminal uptake from typical diets. Calculation of LA balance of the

lactating dairy cow would be the following: LA absorbed from the diet LA used for

maintenance LA used for milk production. Using the fat sub-model of the CPM-Dairy

model, a cow consuming 25 kg of DM/d of a typical diet (no added fat source) would

consume 225 g/d LA. Of the 225 g consumed, only 20% will escape biohydrogenation

(45 g), and of that, 82% (37 g) will be absorbed in the small intestine. The LA

requirement for maintenance of the mature lactating ruminant has not been defined.

However, a calculation based on metabolic body weight using the nonlactating rat

(Mattos and Palmquist, 1977) yields a maintenance requirement of 10.7 g/d for a cow

weighing 607 kg. Milk output of LA of a cow producing 45 kg milk/d would be 54 g/d

(3.4% milk fat containing 3.5% LA). In this situation, the LA balance would be -27.7 g/d

(37 g/d absorbed from the diet 10.7 g/d for maintenance 54 g/d for milk production).

To get this animal out of a deficient situation, a fat rich in LA must be supplemented. A

possible explanation as to why numerous studies report an improvement in reproduction

when additional fat is fed may be due to alleviating a LA deficiency of the modern high-

producing dairy cow.

Many studies report an improvement in reproductive performance of cows fed

supplemental fat. In a review, Staples et al. (1998) reported an improvement in fertility

rates in 11 of 20 articles and speculated that it was a result of dietary FA and not solely









due to an improvement in the energy status of the cows. Although reproductive

performance is strongly associated with energy status (Staples et al., 1990), dietary fats

can provide FA precursors for steroid (including cholesterol) and eicosanoid (including

prostaglandins) production which have an affect on ovarian function, uterine function,

and conception rates.

Dietary fats typically increase concentrations of circulating cholesterol, the

precursor of progesterone (Grummer and Carroll, 1991). Ruminants fed supplemental fat

often have a slight increase in blood progesterone concentration (Staples et al., 1998).

Progesterone, secreted by the corpus luteum (CL), prepares the uterus for implantation of

the embryo and helps maintain pregnancy by providing nourishment for the concepts via

induction of heterotrophic proteins from the endometrium. Work by Hawkins et al.

(1995) suggests that the increase seen in circulating progesterone when cows are fed

supplemental fat is from a reduced rate of clearance of progesterone rather than an

increase in progesterone synthesis. Son et al. (1996) reported greater blood cholesterol

and peak plasma progesterone concentration during the second ovulatory cycle in cows

fed tallow at 2 vs. 0% of dietary DM. Workers at the University of Florida (Garcia-

Bojalil et al., 1998) reported that accumulated plasma progesterone from 0 to 50 DIM

was greater, pregnancy rates improved, and energy status did not change when cows were

fed diets of 2.2% CSFA compared to non fat-supplemented cows.

Through a series of desaturases and elongases, LA (C18:2) can form dihomo-k-

linolenic acid, a direct precursor to the series 1 prostaglandins, or can be further

desaturated to arachidonic acid (C20:4), a direct precursor to the 2 series prostaglandins.

Prostaglandin F2a (PGF2,), synthesized by endometrial tissue, is an important regulator









of parturition and the estrous cycle by causing regression of the CL. Upon conception, it

is important to keep the CL from regressing in order to prevent early embryonic death.

Immediately postpartum, 13, 14-dihydro-15-keto-PGF2, metabolite (PGFM) is important

in regressing the CL of pregnancy. If LA is supplemented in the diet prepartum, more

arachidonic acid may be synthesized leading to higher concentrations of the series 2

prostaglandins and possibly a healthier uterine environment. Alternately, if excess LA is

consumed, it can be converted to eicosadienoic acid (C20:2) instead of arachidonic acid

(Kanduce et al., 1982), increasing the synthesis of the series 3 prostaglandins at the

expense of the series 1 and 2 prostaglandins. It is thought that LA can compete with

arachidonic acid for binding sites of a key enzyme, cyclooxygenase 2 (PGHS-2), that is

necessary for the synthesis of PGF2,. The amount of a particular FA (e.g. LA) stored in

the target tissue may control whether there is an inhibition or stimulation of prostaglandin

synthesis. Reducing PGF2 secretion through dietary fats could improve pregnancy rates

by reducing early embryonic loss around the time of embryo recognition.

Plasma IGF-I concentrations are correlated positively with body condition and

DMI. Low IGF-I concentrations are associated with an extended postpartum interval to

estrus in beef cows and also with delayed puberty (Roberts et al., 1997; Rutter et al.,

1989), indicating that IGF-I can be positively correlated with reproductive performance.

Beam and Butler (1998) reported lower mean concentrations of plasma IGF-I from wk 1

to 3 postpartum in cows fed a diet containing 2.6% prilled fat compared to controls (37.6

vs. 47.7 ng/ml) despite no differences in energy balance. However, other studies reported

no differences in concentration of plasma IGF-I when supplemental fat was fed (Salado

et al., 2004; Spicer et al., 1993). Insulin-like growth factor I acts synergistically with









luteinizing hormone (LH) to promote follicular development (Lucy, 2001). However, if

IGF-I is over stimulated there may be deleterious effects on embryo development, the

uterine environment, and gene expression (Bilby et al., 2004). More specifically,

overstimulation of IGF-I is detrimental to follicle and oocyte development (Armstrong et

al., 2001).

Although the results are somewhat mixed, improvement in conception rates when

fat is supplemented in the diet is often reported. In a study conducted in Wisconsin

(Scott et al., 1995), five herds (n = 443) were fed CSFA at 0 or 450 g/d from 1 tol80 or

200 DIM. They reported an increase in overall conception rate from 93 to 98%, and a

tendency for more fat-supplemented cows to exhibit standing estrus (71.4 vs. 65.6%). In

addition, they reported a tendency for less incidence of noncyclic ovaries in fat-fed cows.

A study conducted in Pennsylvania and Israel by Ferguson et al. (1990) reported an

improvement in first service conception rate when 253 cows over four herds were fed 0

or 2% ruminally inert fat from 0 tol50 DIM (43 vs. 59%). Multiparous Holstein cows (n

= 81) were fed isoenergetic diets containing 1.7% supplemental fat (prilled long chain

FA) for 21 d prepartum and control or glucogenic-supplemented diets for 28 d

postpartum (Frajblat and Butler, 2003). Fat supplementation prepartum did not affect

follicle dynamics measured by ultrasonagraphy nor concentration of plasma

progesterone, insulin, IGF-I, or NEFA. However, supplemental fat prepartum was

associated with better pregnancy rates (86 vs. 58% for fat-supplemented and control cows

respectively, P = 0.03). Recently, workers in Missouri (Oelrichs et al., 2004) reported no

benefit for conception rates of Holstein cows (n = 64) fed raw, cracked soybeans

beginning at 28 d prepartum or beginning at calving (fed at 1.9 and 2.9 kg of DM during









the prepartum and postpartum periods, respectively) despite an improvement in energy

balance. Concentrations of plasma progesterone and PGFM, interval to first estrous

cycle, and rates of cyclicity, ovulation, conception and pregnancy were not different from

cows not fed soybeans. However, cows fed soybeans beginning either prepartum or at

calving had fewer small (< 5 mm) follicles and tended to have more medium (6 to 9 mm)

follicles than controls during the first synchronized estrous cycle. The high LNA and LA

content in flaxseeds (57% LNA and 14% LA) may have been responsible for the

improvement in conception rates (87.5 vs. 50.0%) of lactating dairy cows fed

formaldehyde-treated whole flaxseed (17% of dietary DM) compared to those fed Ca

salts of palm oil (5.6% of dietary DM) from 9 to 19 wk postpartum (Petit et al., 2001).

To investigate the theory that specific FA (e.g., LA) reaching target tissues could

improve conception rates, Santos and coworkers (2004) supplemented dairy cows with a

ruminally inert blend of LA and monoenoic trans FA or a Ca salt of palm oil from 25 d

prepartum through -55 d postpartum when cows were timed AI, then flushed 5 d after AI

and recovered structures were evaluated. Cows fed the LA and monoenoic trans FA

tended to have (P = 0.11) a greater fertilization rate (87 vs. 73%), had more accessory

sperm per structure collected (34 vs. 21), and tended to have (P = 0.06) a greater

proportion of embryos classified as high quality (73 vs. 51%). In an accompanying

study, conception rate at first AI was greater for cows fed the blend of LA and

monoenoic trans FA salt (38.9 vs. 25.9%).

In contrast, a few studies have reported a significant decrease in conception rates of

cows fed supplemental fat. In reviewing three studies that reported decreased conception

rates, Staples et al. (1998) noted that in all studies there was a dramatic improvement in









milk production. High milk production and negative energy balance have been linked to

decreased fertility in dairy cattle.

Effects of Supplemental Fat on Follicle Development

In addition to an improvement in conception rates of lactating dairy cows, follicular

development is improved often by fat feeding. Cows in negative energy balance or in

poor body condition can experience reduced ovarian activity (Staples et al., 1990) which

might be alleviated quicker by supplementing with fats. The mechanism by which

ovarian activity is affected by energy status is likely at the hypothalamus-pituitary axis

and perhaps at the ovary itself. Leutinizing hormone and follicle stimulating hormone

(FSH) are secreted by the anterior pituitary gland upon stimulation by gonadotropin

releasing hormone (GnRH) released from the hypothalamus to cause recruitment and

growth of ovarian follicles. In early lactation and during the state of negative energy

balance, ovarian activity is reduced by low pulsatile secretion of LH (Beam and Butler,

1999).

Simmetal cows (n = 12) were assigned to receive CSFA (0.5% of body weight) or

an isocaloric control supplement in addition to prairie hay from parturition until the

second postpartum ovulation. Calves were permanently removed at 25 d postpartum to

assist with a quicker return to estrus. Concentrations of mean serum LH and total

cholesterol for fat-supplemented cows was greater than for control animals. In addition,

follicular development as determined by ultrasonography was affected in that growth of

class 2 (6 to 9 mm) follicles into class 3 (10 to 15 mm) and 4 (> 15 mm) follicles was

enhanced in cows receiving CSFA (Hightshoe et al., 1991).

At parturition, 18 cows were assigned to receive CSFA at 0 or 2.2% of dietary DM

in a TMR containing 14.5% whole cottonseeds until 60 d postpartum (Lucy et al., 1991).









Prior to 25 d postpartum, CSFA-supplemented cows had a decreased number of 3 to 5

mm follicles and an increased number of 6 to 9 mm follicles. After d 25 postpartum,

estrous was synchronized. The number of 3 to 5 mm follicles and follicles > 25 mm

increased in CSFA-fed cows. In addition, the diameters of the largest and second largest

follicles were greater in CSFA-supplemented cows.

Eighteen lactating Holstein cows were fed CSFA at 2.2% of dietary DM or an

isoenergetic diet (Lucy et al., 1993). Although animals were in similar energy balance,

cows fed CSFA had a larger second wave dominant follicle (18.7 mm) than did cows fed

the 0% CSFA diet (16.1 mm).

Forty-five Holstein cows were fed a blend of tallow and yellow grease (88:12

wt/wt) at 0, 2.2, or 4.4% of the dietary DM from d 0 to 84 DIM (Beam and Butler, 1997).

On d 14 postpartum, the number of follicles greater than 15 mm in diameter was

dramatically increased in cows fed diets of 2.2 and 4.4% (- 0.7) supplemental fat in

comparison to the control (- 0.3) and was not correlated with energy status. The

diameter of the largest follicle from d 8 to 14 postpartum was greater in cows fed 2.2%

supplemental fat (13.5 mm) versus controls (11.0 mm). If only the animals that ovulated

their first wave dominant follicle were considered, all fat-supplemented cows increased

the diameter of the largest follicle from d 8 to 14 postpartum.

At parturition, Holstein cows (n = 141) were allotted to one of three dietary

treatments (Petit and Twagiramungu, 2002). The isonitrogenous, isoenergetic, and

isolipidic diets contained whole flaxseed, Ca salts of palm oil, or micronized soybeans.

The diameter of the CL of the cows fed flaxseed was larger than that of cows fed

soybeans (19.7 vs. 16.9 mm) but not larger than that of cows fed Ca salts of palm oil (17.5









mm). Embryo mortality from day 30 to 50 after AI tended to be lower (P < 0.11) when

cows were fed flaxseed (0%) compared to Ca salts of palm oil (15.4%) or soybeans

(13.6%).

The timing of initiation of fat supplementation during the periparturient period has

received little attention. The beneficial results seen in various studies when feeding fat is

initiated during the periparturient period is not consistent. There is only one published

study (Salfer et al., 1995) conducted to evaluate the timing of initiation of fat

supplementation in lactating dairy cows. The authors concluded that delaying the

inclusion of partially hydrogenated tallow in the diet until 35 DIM had benefits on total

milk production through improved persistency. The objective of the current experiment

was to evaluate if initiating fat supplementation during the prepartum period, at

parturition, or at 28 DIM would have a beneficial effect on milk production, liver

function, and reproduction of Holstein cows during summer.


















Table 2.1. Effects of feeding ruminally inert fat (Ca salts of fatty acids (CSFA) or prilled fat (PF)) to the same cows throughout the

study on DMI, milk production, milk composition, and body weight (BW) change.


Treatments/ Fat source


Dietary
EE,


Erickson et al., 1) Control
1992 2) 12 g/d niacin (NA)
3) 3% CSFA
4) NA + CSFA


Moallem et al., 1) Control
2000 2) 0.55 kg/d CSFA
3) Control + bST


Firkins and
Eastridge, 1992


1) Control
2) 7% soy hulls (SH)
3) 7% SH + 1% NaHCO3
4) 20% SH + 0.43%
CSFA+ 10.7%
roasted soybeans


Garcia-Bojalil 1) 11.1% RDP
et al., 1998 2) 11.1% RDP
CSFA
3) 15.7% RDP
4) 15.7% RDP
CSFA


Spicer et al., 1) Control
1993 2) 1.8% CSFA


1)2.39
2)2.39
3)2.34
4)3.42
(FA)




1)4.77
2)6.65
3)4.62
4) 6.20


DIM


--- 15-98


--- 0-150


28-133


0-120


--- 28-84


Diet, % of DM


35% alfalfa haylage,
10% corn silage,
55% concentrate



9.5% wheat silage,
15% corn silage,
3.5% legume hay,
3.5% oat hay, 2.3%
wheat straw, 66.2%
concentrate

Trt 1 and 4: 10%
alfalfa silage, 31%
corn silage, 59%
concentrate
Trt 2 and 3: 10%
alfalfa silage, 20%
corn silage, 70%
concentrate

34% corn silage,
13% alfalfa hay, 53
% concentrate a


20% sorghum silage,
19% alfalfa hay, 61%
concentrate a


DMI, Milk,
kg/d kg/d


1) 18.5
2) 18.5
3) 17.5
4) 18.2


1)23.5
2)23.7
3)24.5


1)24.4
2)23.5
3)23.2
4)22.5


1) 19.6
2) 19.0
3) 19.4
4) 19.8
Fx RDP
** at 0-50
DIM


1)25.9
2) 24.4


1) 36.2
2) 36.4
3) 38.2
4) 39.3
F**

1) 40.2
2) 42.4
3) 45.4
F**



1) 37.0
2) 37.3
3)35.1
4) 38.9


1)27.1
2) 28.0
3)25.5
4) 27.7
F** at 50-
120 DIM


1) 36.9
2) 36.0


Reference


Milk
protein,
%
1)2.71
2)2.84
3)2.55
4)2.68
F**

1)2.92
2)2.92
3)2.94


4.0%
FCM,
kg/d
1) 32.4
2) 32.7
3) 34.3
4) 35.4
F**

1)37.8
2) 40.8
3) 43.2
t
F**


1)33.8
2) 33.0
3) 31.2
4) 35.6
2 & 3 vs.
4*



1) 25.5
2) 26.5
3)24.1
4) 26.3




1)34.1
2) 32.7


1)3.06
2)3.02
3)3.06
4)2.98
F** at 35-
120 DIM


Milk fat, BW change,
% kg or kg/d

1)3.32 1)0.18
2)3.32 2)0.11
3)3.36 3)-0.22
4)3.35 4)0.07
(kg/d)

1)3.12 N.S.
2)3.25
3)3.19
F**


1)3.50
2)3.26
3)3.39
4)3.42
2 & 3 vs. 4*


1)3.63
2)3.67
3)3.66
4)3.68




1)3.54
2)3.44


1)0.38
2) 0.43
3)0.28
4)0.18
(kg/d) tQ
-4


N.S.


1) 1.4
2)6.3
F**


t 3.5% FCM.
aConcentrate mix included 6-15% whole cottonseed.
F = effect of fat.
N.S. = not significant.
*P <.10.
** P <.05.


1)3.16
2)3.18
3)3.18
4)2.88
F*


















Table 2.1. Continued.
Reference Treatments/ Fat source


Chouinard et 1) Control + NaHCO3
al., 1997 2) 2% CSFA + NaHCO3
3) 4% CSFA + NaHCO3
4) 4% CSFA



Chouinard et 1) Control + NaHCO3
al., 1997 2) 2% CSFA + NaHCO3
3) 4% CSFA + NaHCO3
4) 4% CSFA



Harrison et al., 1) Control
1995 2) 12% WCS
3) 12% WCS and 2.7%
CSFA

Harrison et al., 1) Control
1995 2) 12% WCS
3) 12% WCS and 5%
CSFA


Holter et al., 1) Control
1992 2) 15% WCS
3) 15% WCS + 0.54 kg/d
CSFA


Dietary
EE,
%
1)3.9
2) 5.5
3)6.6
4)6.9


1)4.0
2)5.8
3)7.5
4)7.5



1)2.5
2)4.4
3)6.0



1)3.5
2)5.1
3)6.9


39-67


68-95






21-119





18-105


--- 0-112


Diet, % of DM


62% alfalfa silage,
38% concentrate


46% alfalfa silage,
54% concentrate





23% alfalfa hay, 23%
grass silage, 54%
concentrate



28% alfalfa hay, 18%
grass silage, 54%
concentrate



Ad libitum forage:
63% corn silage,
37% wilted grass
silage. Concentrate
was adjusted to milk
production


DMI, Milk, 4.0%
kg/d kg/d FCM,
kg/d
1)23.6 1)33.5 1)31.2
2)22.8 2)35.2 2)31.6
3)21.4 3)32.8 3)28.9
4)21.6 4)32.0 4)28.0
F** F** t F**
(linear) (quadratic) (linear)


1)23.4
2)21.9
3)21.5
4)21.0



1)23.1
2)23.9
3)21.6
Trt**

1)23.6
2)22.4
3)22.2
Trt**
Trt xP**

1) 17.4
2) 16.6
3) 16.8


1)33.6
2)35.9
3) 34.0
4)31.4
Fx
NaHCO3*


1)35.2
2)29.6
3)32.5
Trt**


1) 30.4
2) 31.3
3) 28.7
4) 26.7



1)38.1
2)39.8
3)39.5
Trt**

1)36.0
2)37.2
3)37.8
Trt**
Trt x P**


1)31.6
2)30.3
3)31.9


Milk
protein,
%
1)2.96
2)2.80
3)2.66
4)2.57
F**


1)3.10
2)2.96
3)2.82
4) 2.74
F**

1)3.08
2)3.07
3)2.91
Trt**

1)3.09
2)3.11
3)2.95
Trt**
Trt xP**

1)2.86
2)2.88
3)2.82
Trt**


Milk fat, BW change,
% kg or kg/d


1)3.55
2)3.38
3)3.29
4)3.43


1)3.41
2)3.08
3)2.93
4)3.23
F** linear

1)3.24
2)3.49
3)3.74
Trt**

1)3.36
2)3.65
3)3.72
Trt**
Trt xP**


1)3.32
2)4.14
3)3.89
Trt**


t 3.5% FCM.
WCS = whole cottonseed.
F = effect of fat.
Trt = effect of treatment.
P = effect of parity.
N.S. =not significant.
*P <.10.
** P <.05.


















Table 2.1. Continued.
Reference Treatments/ Fat source


Selberg et al., 1) Control
2004 2) 225 g/d CaS
conjugated linoleic
acid
3) 225 g/d CaS trans-
C18:1



Hoffman et al., 1)Degradable protein
1991 (DP)
2) Undegradable protein
(UP)
3) DP + 2.8% sodium
alginate treated tallow
(SAT)
4) UP + 2.8% SAT

Beam and 1) Control
Butler, 1998 2) 2.59% PF






Jerred et al., 1) Low silage (LS)
1990 b 2) LS + 5% PF
3) Medium silage (MS)
4) MS + 5% PF
5) High silage (HS)
6) HS + 5% PF


Dietary
EE,
%0
Prepartum
control:
4.3
Post-
partum
control:
5.2


1)3.1
2) 3.3
3)5.7
4)6.0






1)4.8
2)7.0






1)3.1
2)6.5
3)3.4
4)7.2
5)3.8
6)7.2


DIM


-28-49


Diet, % of DM


Prepartum: 13%
bermuda grass hay, 39%
corn silage, 52%
concentrate
Postpartum: 10% alfalfa
hay, 29% corn silage,
61% concentrate


22-150 49% alfalfa silage, 51%
concentrate









0-100 26% corn silage, 18%
alfalfa haylage, 56%
concentrate




5-105 Trt 1 & 2: 45% alfalfa
silage, 55% concentrate
Trt 3 & 4: 64% alfalfa
silage, 36% concentrate
Trt 5 & 6; 84% alfalfa
silage, 16% concentrate


DMI, Milk,
kg/d kg/d


1)21.6
2)20.0
3)20.2
1 vs. 3 at
wk 4-6**
1 vs. 2 at
wk6**


1)22.7
2) 22.5
3)22.7
4) 22.7
(group
fed)




0-100
DIM N.S.
0-28 DIM
1) 15.5
2) 17.3
F**

1)23.6
2)22.1
F**
Fx T**


1) 40.3
2)41.5
3)41.5
1 & 2 vs. 3
by wk**




1)31.8
2)31.6
3)33.1
4) 32.7
FxT**





FxT*
(Control
peaked -41
vs. 43 kg/d
for PF)


1) 39.2
2)38.8


4.0% FCM, Milk
kg/d protein,
%


1) 40.0
2) 37.4
3) 40.5
T


1) 29.9
2) 29.9
3) 30.9
4) 31.2


1)2.89
2) 2.82
3)2.81


1)3.14
2) 3.03
3) 3.04
4) 3.00
F**
FxT**


FxT*
(Control
peaked -34
vs. 36 kg/d
for PF)


1)36.5
2) 37.8
FxT**


1)2.89
2) 2.87
FxT**


t 3.5% FCM.
aConcentrate mix included 6-15% whole cottonseed.
b Note: there are no fat x forage interactions; results are presented as 1) control (treatments 1, 3 and 5) and 2) fat (treatments 2, 4, and 6).
F = effect of fat.
T effect of time.
N.S. =not significant.
*P <.10.
** P <.05.


Milk fat,
%

1)3.49
2)2.99
3)3.46
1 vs. 2**


1)3.67
2) 3.63
3)3.60
4)3.82


BW change,
kg or kg/d


N.S.










1)-26.5
2)-42.6
F*





1) -0.36
2) -0.47
(kg/d)


1)3.57
2)3.88
F**


















Table 2.1. Continued.
Reference Treatments/ Fat source


Skaar et al., 1) Control
1989 2) 12 g/d niacin (NA)
3) 5% PF
4) 12 g/d NA and 5% PF





Pickett et al., 1) Control
2003 2) 500 ml/d propylene
glycol (PG) drench
3) 0.45 kg/d CSFA
drench
4) 500 ml/d PG + 0.45
kg/d CSFA drench


Schroeder et al.,
2003


1) TMR fed (control)
2) Pasture + 6.7 kg/d corn
based concentrate
3) Pasture + 6.7 kg/d
concentrate with 0.8 kg
CSFA


Dietary
EE,
%


Diet, % of DM


1)3.4b -17-105 25% corn silage, 25%
2)3.4 b alfalfa silage, 50%
3) 11.8b concentrate
4) 11.8b





4.8 0-21 31% corn silage, 16%
alfalfa hay, 9% alfalfa
hay, 44% concentrate
Drenches were
administered from 0-3
DIM


1)4.5
2)6.1
3)8.1


Moallem et al., 1) Control
1999 2) 0.55 kg/d CSFA
3) Control + bST


Kim et al., 1993


1) Control
2) 17% extruded
soybeans
3) 4.0% CSFA


117-152 Control: 59% corn
silage, 41% concentrate
Treatments 2 and 3:
84% pasture, 16%
concentrate


--- 0-150 8% wheat silage, 20%
corn silage, 3% pea hay,
3% oat hay, 66%
concentrate a


1)2.5
2)5.1
3)3.0


28-105 25' i11i .i 1 25%
corn silage, 50%
concentrate


DMI, Milk,
kg/d kg/d


1) 19.3
2) 17.8
3) 19.2
4) 18.7





1) 17.2
2) 18.0
3) 16.9
4) 15.8




1)23.7
2)22.9
3)21.5


1)24.0
2)23.3
3)24.7
(group
fed)
1) 17.8
2) 18.4
3) 16.6
2 vs. 3*


1) 38.4
2) 36.3
3) 42.0
4)41.3
F x season**
(fat greater
in summer)

1)36.5
2)36.1
3)32.5
4) 34.8


1) 20.2
2) 19.2
3) 20.2


1) 39.7
2) 42.5
3) 44.0
F**

1)29.2
2)32.4
3)31.8
F**


4.0% FCM, Milk
kg/d protein,
%


1) 36.3
2) 34.5
3) 39.3
4) 38.2
T Fx
season** (fat
was greater
in summer)
1) 42.9
2) 40.3
3)38.1
4) 40.9
1t


1) 19.5
2) 17.8
3) 16.1
1 vs. 3**



1)37.8
2) 40.9
3) 41.9
t
F**
1)25.4
2) 26.7
3) 28.7
F*


1) 3.00
2) 2.87
3) 2.87
4) 2.87


1) 3.68
2) 3.46
3) 3.66
4)3.51


1) 3.70
2) 3.49
3)3.41
1 vs. 2 &
3**
Trt x T**


1) 2.98
2) 2.92
3) 2.92
F**

1)2.99
2)2.93
3)2.81
F**
2 vs. 3**


t 3.5% FCM.
aConcentrate mix included 6-15% whole cottonseed.
b concentrate mix only.
F = effect of fat.
T effect of time.
Trt = effect of treatment.
N.S. =not significant.
*P <.10.
** P .05.


Milk fat,
%

1)3.14
2)3.19
3)3.15
4)3.12


BW change,
kg or kg/d

FxT**
(fat fed
gained
faster)


1)4.66
2)4.30
3)4.64
4) 4.66


1)23
2)-6
3)-10
1 vs. 2 &
3**


Fat fed lost
more BW
(-7 kg) than
control (P<
0.1)


1)3.91
2)3.45
3)2.56
Trt**
Trt x T**


1)3.18
2)3.25
3)3.19


1)3.20
2)2.69
3)3.47
2 vs. 3**


















Table 2.1. Continued.
Reference Treatments/ Fat source


Moallem et al., 1) Control
1997 2) 0.5 kg/d CSFA
3) Control + bST
4) 0.5 kg/d CSFA + bST


Sklan et al., 1) Control
1994 2) 2.5% CSFA


Cervantes et al.,
1996


1) Control
2) 0.4 kg/d CSFA
3) 12 g/d nicotinamide
(NM)
4) 0.4 kg/d CSFA + 12
g/d NM


Palmquist and 1) Control
Weiss, 1994 2) 2.5% tallow + 2.5%
CSFA
(Also included 3
concentrations of
RUP)

Atwal et al,, 1) Control
1990 2) 5% CSFA


Dietary
EE,


Diet, % of DM


--- 0-150 10.4% wheat silage,
24% corn silage, 2.3%
pea hay, 2.1% oat hay,
61.2% concentrate


1)2.8
2)4.9


1)3.1
2)5.1
3)3.0
4)5.0


1)3.08
2) 6.73
(FA)


0-120 14.3% wheat silage,
15.3% corn silage, 6.4%
vetch hay, 64%
concentrate a


112-150 Varied dependent on
stage of lactation.
Forages (35-60% of
diet) utilized were
i i ,I alfalfa
haylage, and corn
silage.


0-60


25% corn silage, 25%
alfalfa hay, 50%
concentrate


1)2.4 14-70 15% alfalfa silage, 10%
alfalfa hay, 25% corn
silage, 50% concentrate


DMI, Milk,
kg/d kg/d


1)24.1
2) 24.3
3)24.9
4) 24.7
(group
fed)


1)20.6
2)20.3
(group
fed)


1)20.3
2)20.5
3)24.0
4)21.1
FxNM*


1) 36.3
2) 39.8
3) 41.3
4) 42.9
F**
Fx bST**


1)31.1
2)35.0
F**
FxT**


1)30.7
2)31.8
3)33.5
4)33.2


1) 18.1 1)43.3
2) 18.8 2)42.4


1) 19.2
2) 19.7


1)33.1
2)33.3


4.0% FCM, Milk
kg/d protein,
%


1)33.4
2) 36.9
3)39.5
4) 40.8
F**
Fx bST**


1) 29.4
2)33.8

F**
FxT**

1) 28.3
2) 29.7
3) 29.7
4) 30.6


1)37.1
2)37.1





1)31.1
2)32.5


1) 3.02
2) 2.99
3) 3.01
4) 2.99




1) 2.97
2) 2.99
FxT**



1) 3.21
2)3.17
3)3.31
4)3.14
F**


Milk fat,
%

1)2.98
2)3.03
3)3.21
4)3.20




1)3.15
2)3.26
F*
FxT**


1)3.45
2)3.57
3)3.26
4)3.46
F*


1)2.97 1)2.64
2)2.93 2)2.78


1)2.93 1)3.51
2)2.94 2)3.80


t 3.5% FCM.
aConcentrate mix included 6-15% whole cottonseed.
F = effect of fat.
T effect of time.
*P <.10.
**P<.05.


BW change,
kg or kg/d

1) 38.0
2) 43.2
3)33.4
4) 35.4
(maximum
BW loss)


1)-0.08
2)-0.23
(kg/d)


1)0.10
2) 1.35
(kg/d)
F** (wk 1-4)


















Table 2.1. Continued.
Reference Treatments/ Fat source


Atwal et al,, 1) Control
1990 2) 5% CSFA


Simas et al., 1) Dry rolled sorghum
1995 (DRS)
2) Steam flaked sorghum
(SFS)
3) DRS + 2.5% CSFA
4) SFS + 2.5% CSFA


Sklan et al., 1) Control
1991 2) 2.6% CSFA


Sklan et al., 1) Control
1992 2) 2% FA
3) 2.4% CSFA




Sklan et al., 1) 14.5% WCS
1992 2) 1.8% FA
3) 2.1% CSFA


Dietary
EE,


1)2.3 1-56


1) 5.7
2)5.7
3)7.5
4)7.5


Diet, % of DM


25' 11 1 1. 1 25%
corn silage, 50%
concentrate


34 ,11 ,11 66%
concentrate'


--- 0-120 13.7% corn silage,
11.3% vetch hay, 75%
concentrate


1)2.1
2)4.2
3)4.2
(FA)



1) 5.0
2)4.0
3)4.0
(FA)


93-213


1.4 ...1 20%
wheat silage, 13.7%
corn cobs, 64.9%
concentrate


108-228 7.8% citrus silage,
32.7% wheat silage,
6.7% vetch hay,
52.8% concentrate


DMI, Milk,
kg/d kg/d

1) 16.5 1)30.1
2) 15.1 2)29.7


1)21.6
2)23.1
3) 17.8
4) 19.5
F**



1)20.3
2)20.2
(group
fed)


1)21.2
2)21.1
3)20.9
(group
fed)

1)20.1
2)20.4
3)20.3
(group
fed)


1) 34.3
2) 39.3
3)33.4
4) 36.5


1) -37
2) -39
(at peak)
F** at 30
and 60 DIM


1) 30.2b
2) 31.3a
3) 30.9ab
Trt xT**
Trt xP**


1) 31.3b
2) 32.5a
3) 32.4ab
Trt xT**


4.0% FCM, Milk
kg/d protein,
%


1) 32.6
2) 30.8



1) 31.6
2) 34.5
3) 31.2
4) 32.8
T


1) -34
2) -38
(at peak)
F** until 90
DIM


1) 25.9c
2) 27.7b
3) 27.0a
T
Trt x T**
Trt xP**
1)30.1
2) 29.7
3) 30.4

Trt xT
Trt x T*


Milk fat,
%


1)3.03 1)4.82
2) 3.01 2)4.37


1) 2.93
2) 3.00
3) 2.79
4) 2.99
3 vs.
others*


1)3.16
2)2.91
3)3.23
4)3.05


N.S. 1)-2.8
2) -3.0
F** at 30 to 90
DIM


1)3.20
2)3.13
3)3.15
Trt xT**
Trt xP**


1)3.02
2)2.96
3)2.98
Trt xT*


1)2.67a
2) 2.81b
3) 2.75ab
Trt xT**
Trt xP**


1)3.25a
2) 2.99b
3)3.17a


t 3.5% FCM.
a,b,c Means not followed by the same letter differ (P < .05).
1 Concentrate mix included 6-15% whole cottonseed (WCS).
F = effect of fat.
Trt = effect of treatment.
T effect of time.
P = effect of parity.
N.S. =not significant.
*P <.10.
** P <.05.


BW change,
kg or kg/d

1)-0.35
2)-1.85
(kg/d)


1)0
2)0.16
3) -0.2
4) -0.02
(kg/d)
F*


F**
(CSFA cows
lost more
and faster)


















Table 2.2. Effects of feeding oilseeds alone or in combination with other fat sources to the same cows throughout the experiment on

DMI, milk production, milk composition, and body weight (BW) change.


Reference


Stegeman et al.,
1992


AbuGhazaleh et
al., 2004



Kim et al., 1993


Treatments/ Fat source


1) Control
2) Control + bST
3) 10% rolled sunflower
seeds + bST
4) 10% rolled safflower
seeds+ bST


1) Control
2) 10.64% extruded
soybeans + 5.5% fish
meal

1) Control
2) 17% extruded
soybeans
3) 4.0% CSFA


Markus et al., 1) Control
1996 2) 7.1% whole sunflower
seeds
3) 2.7% tallow

Weiss and 1) Control
Wyatt, 2003 2) 12.3% whole roasted
soybeans
3) 2.35% tallow
(Also included 3 levels of
vitamin E)
t 3.5% FCM.
F = effect of fat.
T effect of time.
P<.10.
** P <.05.


Dietary
EE,
%
1)2.3
2)2.3
3)6.2
4) 5.5




1)2.67
2) 4.93



1)2.5
2)5.1
3)3.0


1) 1.8
2)4.2
3)4.1


DIM


Diet, % of DM


104-216 25% corn silage,
25% alfalfa hay, 50%
concentrate


103-173 25% alfalfa hay, 25%
corn silage, 50%
concentrate


28-105


16-112


25% alfalfa hay, 25%
corn silage, 50%
concentrate



12% corn silage,
14% alfalfa silage,
9.5% alfalfa hay,
64.5% concentrate


--- 160-188 38% corn silage, 8%
alfalfa hay, 7%
alfalfa silage, 47%
concentrate


DMI, Milk,
kg/d kg/d


1)27.5
2)23.9
3)27.3
4) 19.3
3 vs. 4**



1)29.3
2) 27.7
F*


1) 17.8
2) 18.4
3) 16.6
2 vs. 3*


1)22.2
2)21.1
3)21.6


1)22.3
2) 24.0
3)22.0
2 vs. 3**


1)29.5
2) 32.7
3) 40.0
4)34.1
2 vs. 3 &
4**
3 vs. 4**

1)34.5
2) 38.9
F**
FxT**

1) 29.2
2) 32.4
3)31.8
F**


1)34.4
2)34.6
3)35.5


1)35.1
2)36.8
3)37.5
F*


4.0%
FCM,
kg/d
1)25.0
2) 27.9
3) 32.3
4)28.1
3 vs. 4**



1) 36.0
2) 36.4
t


1)25.4
2) 26.7
3) 28.7
F*


1)30.0
2)29.9
3)31.6


Milk
protein,
%
1)3.17
2)3.27
3)3.02
4)3.08
F**
2 vs. 3 &
4**

1)3.39
2)3.18
F**
Fx T**

1)2.99
2)2.93
3)2.81
F**
2 vs. 3**

1)3.1
2)3.0
3)3.0


1)2.97
2)2.92
3)2.86


Milk fat, BW change, kg
% or kg/d

1)2.99 1)24.7
2)3.06 2)19.4
3)2.73 3)36.1
4)2.86 4)39.9


1)3.74
2)3.17
F**
Fx T**

1)3.20
2)2.69
3)3.47
2 vs. 3**


1)3.2
2)3.1
3)3.3


1)3.76
2)3.83
3)3.08
F**
2 vs. 3**


1)0.87
2) 1.29
3)0.71
(kg/d)
2 vs. 3**


















Table 2.2. Continued.
Reference Treatments/ Fat source


Drackley et al., 1) Control
1998 2) Control + 12 g/d niacin
3) 10% whole raw
soybeans and 2.5%
tallow
4) 10% whole raw
soybeans, 2.5% tallow
and 12 g/d niacin

Sklan et al., 1) Control
1992 2) Control + 2.4% FA3
3) Control + 16.2% WCS




Sklan et al., 1) 14.5% WCS
1992 2) 1.8% FA3
3) 2.1% CSFA


Dietary
EE,
%
1)2.75
2)2.75
3)6.04
4) 6.04
(FA)


1) 1.8
2)4.4
3)4.4
(FA)



1) 5.0
2)4.0
3)4.0
(FA)


28-301


70-210


Diet, % of DM


32.5% alfalfa
haylage, 17.5% corn
silage, 50%
concentrate 1 2


8.4% alfalfa hay,
17.2% corn silage,
74.4% concentrate


108-228 7.8% citrus silage,
32.7% wheat silage,
6.7% vetch hay,
52.8% concentrate


DMI, Milk,
kg/d kg/d


1)21.9
2)21.7
3)21.6
4) 22.2
FxT**


1)21.4
2)22.1
3)21.4
(group
fed)

1)20.1
2)20.4
3)20.3
(group
fed)


1)30.5
2) 33.2
3)31.8
4) 33.6
FxT**


1) 32.7
2) 33.6
3)33.5
Trt xT**



1) 31.3b
2) 32.5a
3) 32.4ab
Trt xT**


4.0%
FCM,
kg/d
1) 30.4
2) 32.9
3)32.5
4) 34.2
FT
FxT**


1) 27.6b
2) 28.7a
3) 30.4a

Trt x T**

1)30.1
2) 29.7
3) 30.4
Trt x
Trt xT**


Milk
protein,
%
1)3.29
2)3.16
3)3.16
4)3.13
F**
FxT**




1)2.97
2)2.95
3)2.96
Trt xT*



1)3.02
2)2.96
3)2.98
Trt xT*


Milk fat, BW change, kg
% or kg/d


1)3.56
2)3.50
3)3.68
4)3.60
F x niacin x
T**


1)2.54b
2)2.57b
3) 2.96a
Trt xT*



1)3.25a
2) 2.99b
3)3.17a


1 Concentrate mix included 6-15% whole cottonseed (WCS).
2After 175 DIM, diets were adjusted for decreased nutrient requirements. Forage content increased to 60% of DM.
to 2.25% of the diet.
3 Fatty acid (FA) source was mixed soapstock containing 86% free FA, of which 20% was linoleic acid.
a,b,c Means not followed by the same letter differ (P < .05).
t 3.5% FCM.
F = effect of fat.
Trt = effect of treatment.
T effect of time.
*P <.10
** P <.05


For treatments 3 and 4 whole raw soybeans were removed and tallow was decreased























1) Control
2) Added fat from
extruded soybeans or
sunflower seeds
(summary of 5 studies)


Dietary
EE,
%
1)2.6
2)5.3
(average)


Diet, % of DM


28-119


38% corn silage,
12% alfalfa hay, 50%
concentrate


DMI, Milk,
kg/d kg/d


1)20.8
2)20.4


4.0%
FCM,
kg/d


1) 3000
2) 3085
(kg)


Milk
protein,
%
1)2.99
2)2.91
F**


Milk fat, BW change, kg
% or kg/d

1)3.20 1)0.18
2)2.95 2)0.13
F** (kg/d)


Harrison et al., 1) Control
1995 2) 12% WCS
3) 12% WCS and 2.7%
CSFA


Harrison et al., 1) Control
1995 2) 12% WCS
3) 12% WCS and 5%
CSFA


Khorasani et al.,
1991


1) Control
2) 4.5% jet-sploded
whole canola seed
(JSWCS)
3) 9% JSWCS
4) 13.2% JSWCS
5) 17.4% JSWCS


Faldet and 1) 10% soybean meal
Satter, 1991 2) 13% raw soybeans
(RS)
3) 13% heated soybeans
(HS)
S3.5% FCM.
WCS= whole cottonseeds.
F = effect of fat.
P = effect of parity.
Trt = effect of treatment.
P<.10.
** P <.05.


1)2.5
2)4.4
3)6.0



1)3.5
2)5.1
3)6.9




1)2.2
2)3.4
3)4.4
4) 5.5
5)6.7




1)3.3
2)5.6
3)5.6


21-119






18-105


36-92


15-119


23% alfalfa hay, 23%
grass silage, 54%
concentrate



28% alfalfa hay, 18%
grass silage, 54%
concentrate


30% alfalfa silage,
10% oat silage, 60%
concentrate


50% alfalfa silage,
50% concentrate


Table 2.2. Continued.
Reference Treatments/ Fat source


Schingoethe
and Casper,
1991


1)38.1
2) 39.8
3)39.5
Trt**


1)36.0
2) 37.2
3) 37.8
Trt* *
Trt x P**


1)23.1
2)23.9
3)21.6
Trt**


1)23.6
2)22.4
3)22.2
Trt**
Trt xP**


1) 17.8
2) 18.8
3) 18.3
4) 16.2
5) 17.0




1)23.4
2)22.3
3)23.6


1)3.08
2)3.07
3)2.91
Trt**


1)3.09
2)3.11
3)2.95
Trt**
Trt xP**


1)2.95
2)3.12
3)2.89
4)2.84
5)2.71
F** (linear)


1)2.99
2)2.89
3)2.85
HS**


1)3.24
2)3.49
3)3.74
Trt**


1)3.36
2)3.65
3)3.72
Trt**
Trt xP**


1)3.08
2)2.89
3)3.06
4) 2.76
5)2.84




1)3.41
2)3.50
3)3.41


1)32.5
2)34.5
3)34.0
4)33.2
5)29.8




1)34.5
2) 34.2
3) 38.9
HS**


1)27.7
2)28.2
3)29.8
4) 26.9
5)24.8




1)33.4
2) 34.7
3) 38.0
HS**
HS**















Table 2.2. Continued.
Reference Treatments/ Fat source


Holter et al., 1) Control
1992 2) 15%WCS
3) 15% WCS + 0.54
kg/d CSFA



Wu et al., 1) Control
1994 2) 12% WCS
3) 12% WCS + 2.2%
safflower oil
4) 12% WCS +2.2%
prilled tallow
5) 12% WCS +4.4%
prilled tallow


Dietary
EE,


DIM


--- 0-112


1)3.3
2)5.2
3)7.4
4) 7.4
5) 9.6


Kim et al., 1) Basal containing 1)2.6
1991 14% soybean meal 2)5.5
(SBM) 3)5.1
2) 17% extruded
soybeans (ESB)
3) 17% ESB and 5%
SBM
Pires et al., 1) Control 1)3.2
1996 2) 18% ground roasted 2) 6.2
soybean 3) 6.2
3) 18% whole soybean 4) 3.2
4) 2.7% blood meal 5)6.2
5) 2.7% blood meal
+3% tallow
a,b,c Means not followed by the same letter differ (P < .05).
t 3.5% FCM.
WCS = whole cottonseeds.
Trt = effect of treatment.
*P <.10.
** P<.05.


Diet, % of DM


Ad libitum forage:
63% corn silage,
37% wilted grass
silage. Concentrate
was adjusted to
milk production


50-125 43% alfalfa hay,
57% concentrate







28-112 25% corn silage,
25% alfalfa hay,
50% concentrate




21-126 30% corn silage,
20% alfalfa silage,
50% concentrate


DMI, Milk,
kg/d kg/d


1)17.4
2)16.6
3)16.8


1)28.2
2)27.2
3)28.8
4)26.8
5)24.1




1)20.9
2)20.7
3) 19.8




1) 23.6a
2) 22.7ab
3) 21.3bc
4) 21.3bc
5) 20.4c


1)35.2
2)29.6
3) 32.5
Trt**


1)32.5
2) 32.6
3) 35.0
4) 34.3
5) 33.0
1 & 2 vs. 3
&4**


1)33.0
2)35.8
3) 34.2
F**



1)39.6
2)40.7
3) 36.4
4) 36.1
5) 39.3


4.0%
FCM,
kg/d
1)31.6
2) 30.3
3)31.9


1)32.4
2) 32.3
3) 33.4
4) 34.6
5) 33.0

1 & 2 vs.
3 &4**

1)28.5
2)29.6
3) 30.2




1)35.4
2)35.0
3) 33.3
4)33.9
5) 35.0


Milk
protein,
%
1)2.86
2)2.88
3)2.82
Trt**



1)3.20
2)3.03
3) 3.03
4)3.08
5)3.07
1 vs. 2 **



1)2.92
2)2.88
3)2.83




1) 3.03a
2) 2.83b
3) 2.88bc
4) 3.08a
5) 2.98ac


Milk fat, BW change,
% kg or kg/d

1)3.32 ---
2)4.14
3)3.89
Trt**


1)3.49
2)3.48
3)3.26
4)3.58
5)3.51




1)3.20
2)2.88
3)3.17
2 vs. 3*



1)3.33
2)3.09
3) 3.50
4)3.63
5)3.29


1)0.70
2)0.82
3)0.57
4)0.64
5)0.46
(kg/d)



1)0.31
2)0.00
3) -0.02
(kg/d)

















Table 2.3. Effects of feeding rendered fats to the same cows throughout the study on DMI, milk production, milk composition and

body weight (BW) change.


Reference Treatments/ Fat source


Tackett et al., 1)21% NDF
1996 2) 21% NDF + 6% choice
white grease (CWG)
3) 28% NDF
4) 28% NDF + 6% CWG



Niagono et al., 1) Control
1991 2) Control + 1 kg yellow
grease
(Also included high and
low degradability
protein supplements)

Bateman et al., 1) 33% NDF, 0% tallow
1996 2) 33% NDF, 2% tallow
3) 40% NDF, 0% tallow
4) 40% NDF, 2% tallow


Bateman et al., 1) 33% NDF, 0% tallow
1996 2) 33% NDF, 2% tallow
3) 40% NDF, 0% tallow
4) 40% NDF, 2% tallow


Son et al., 1996 1) 0% tallow + 0% escape
protein supplement
(EP)
2) 0% tallow + 5% EP
3) 3% tallow + 0% EP
4)3% tallow +5% EP
S3.5% FCM
a concentrate mix only.
F = effect of fat.
T effect of time.
*P<.10.
** P <.05.


Dietary
EE,


1)2.8
2) 8.4
3)2.7
4)7.8


1) 5.0a
2) 16.3a


1)3.5
2)5.4
3)3.7
4)5.8


1)3.5
2)5.4
3)3.7
4)5.8


DIM


Diet, % of DM


32-122 Treatment 1 & 2: 36%
corn silage, 9% alfalfa
hay, 55% concentrate
Treatment 3 & 4: 36%
corn silage, 16% alfalfa
hay, 48% concentrate


1-112 Ad libitum wheat silage
and maximum of 16.4
kg/d of concentrate




120-240, 11 1 22%
winter corn silage, 14% alfalfa
silage, 0 or 13% earlage,
56 or 43% concentrate


120-240, S 111 22%
summer corn silage, 14% alfalfa
silage, 0 or 13% earlage,
56 or 43% concentrate


14-84 33% alfalfa haylage,
17% corn silage, 50%
concentrate


DMI, Milk, 4.0% FCM, Milk
kg/d kg/d kg/d protein,
%


1)24.4
2)21.7
3)23.7
4)21.5




1)20.2
2) 19.0


1)25.2
2)22.6
3)23.3
4) 24.3
F x NDF*

1)21.7
2)21.9
3)22.7
4) 22.0


1)25.1
2) 24.2
3)24.1
4)22.8
F**
F x EP**


1)37.5
2) 38.9
3) 34.7
4) 38.0


1)37.0
2)35.4
3)34.8
4)36.3
t


1)38.9 1)30.4
2)39.8 2)29.4


1)31.1
2)31.2
3)28.6
4) 30.3


1)29.4
2)30.1
3)30.7
4) 29.9


1)32.5
2)33.0
3)33.0
4)33.1


1)30.3
2)30.2
3)28.1
4)28.9


1)26.9
2)27.5
3)29.8
4) 27.3


1)31.4
2)30.1
3)31.1
4)31.7
F x EP**


1)3.5
2)3.3
3)3.4
4)3.4
F*



1)3.3
2)3.4


1)3.30
2)3.17
3)3.22
4)3.09


1)3.14
2)3.03
3)3.25
4)3.07


1)2.97
2)2.80
3)2.87
4) 2.75


Milk fat, BW change, kg
% or kg/d

1)3.4 1)55
2)2.9 2) 53
3)3.5 3)42
4) 3.2 4) 29
F**
Fx NDF x
T**


1)2.5
2)2.4


1)3.94
2)3.90
3)3.96
4)3.73


1)3.52
2)3.49
3)3.86
4)3.56


1)3.78
2)3.41
3)3.61
4)3.71


1)-2.6
2) -24.4
F**


1)-30.2
2)-20.3
3)-17.3
4)-37.1
F x EP**


















Table 2.3. Continued.
Reference Treatments/ Fat source


Maiga et al., 1) Control
1995 2) 2% tallow
3) 8.3% tallow-molasses
blend
4) 2% tallow + 5.4%
whey

Pantoja et al., 1) Control
1996 2) 5% tallow
3) 5% tallow + partially
hydrogenated tallow
(PHT) (2:1
weight/weight)
4) 5% tallow + PHT(1:2
weight/weight)
5) 5% PHT

Salfer et al., 1) Control
1995 2)-14-0 DIM 1%
partially hydrogenated
tallow (PHT), 1-151
DIM 2% PHT
3) 1-151 DIM 2% PHT
4) 35-151 DIM 2% PHT



Salfer et al., 1) Control
1995 2)-14-0 DIM 1%
partially hydrogenated
tallow (PHT), 1-151
DIM 2% PHT
3) 1-151 DIM 2% PHT


Dietary
EE,
%
1)2.9
2)4.8
3)4.7
4)4.6


1)2.90
2)6.13
3)6.80
4) 6.58
5)6.54
(FA)





Prepartum
1)3.01
2)3.92

Post-
partum:
1)3.17
2) 4.99


Prepartum
1)3.01
2)3.92

Post-
partum:
1)3.17
2) 4.99


DIM


28-112


28-133










1-151










1-35
(3 treatments
initiated)


Diet,% of DM


25% corn silage,
250 ill ill 1 .
50% concentrate


25% corn silage,
25% alfalfa silage,
50% concentrate








Prepartum:
37% corn silage,
10% alfalfa silage,
14% grass hay,
39% concentrate
Postpartum:
24% corn silage,
21% alfalfa silage,
55% concentrate

Prepartum:
37% corn silage,
10% alfalfa silage,
14% grass hay,
39% concentrate
Postpartum:
24% corn silage,
21% alfalfa silage,
55% concentrate


DMI, Milk,
kg/d kg/d


1)23.1
2) 24.3
3) 24.5
4) 24.5



1) 22.3
2)22.1
3)21.2
4) 22.5
5) 23.9
2 vs. 5*





1) 19.9
2) 19.9
3)20.6
4) 20.6







1) 15.3
2) 15.5
3) 16.3


1)31.9
2)33.7
3)33.7
4) 34.0
F**


1)35.6
2) 40.6
3)36.9
4)39.3
5)38.0
F**





1)30.7
2)31.6
3)31.9
4)32.7







1)30.2
2)29.9
3)30.6


4.0% FCM, Milk
kg/d protein,
%


1)31.4
2)33.9
3)33.0
4)33.4
t


1)33.6
2)35.6
3)33.6
4) 36.3
5)36.3






1)32.0
2)32.7
3)33.2
4)32.8
t





1)34.5
2)33.4
3)36.9
vs. 3*
2 vs. 3*


1)3.00
2)2.98
3)2.91
4)2.86
2 vs. 3 & 4*


1)3.05
2)2.86
3)3.03
4)2.98
5)3.02
2 vs. 5**





1)3.04
2)2.99
3)2.97
4) 2.92







1)3.06
2)3.08
3)3.13


Milk fat, BW change, kg
% or kg/d


1)3.48
2)3.65
3)3.52
4)3.40
2 vs. 3 & 4*


1)3.63
2)3.17
3)3.48
4)3.56
5)3.77
2 vs. 5**





1)3.81
2)3.76
3)3.83
4)3.55







1)4.45
2) 4.29
3)4.81
2 vs. 3**


1)0.75
2) 0.42
3)0.60
4)0.61
(kg/d)
F*

1)-2.16
2)-0.53
3) 1.09
4)0.13
5)0.04
(kg/wk)
F**


t 3.5% FCM.
F = effect of fat.
N.S. =not significant.
*P <.10.
** P <.05.


















Table 2.3. Continued.
Reference Treatments/ Fat source


Salado et al., 1) Control
2004 2) 0.7 kg/d partially
hydrogenated
vegetable oil

Markus et al., 1) Control
1996 2) 7.1% whole sunflower
seeds
3) 2.7% tallow

Weiss and 1) Control
Wyatt, 2003 2) 12.3% whole roasted
soybeans
3) 2.35% tallow
(Also included 3 levels of
vitamin E)
Wu et al., 1994 1) Control
2) 12% WCS
3) 12% WCS + 2.2%
safflower oil
4) 12% WCS +2.2%
prilled tallow
5) 12% WCS +4.4%
prilled tallow


Pires et al., 1) Control
1996 2) 18% ground roasted
soybean
3) 18% whole soybean
4) 2.7% blood meal
5) 2.7% blood meal + 3%
tallow
a Concentrate mix only.
WCS= whole cottonseeds.
F = effect of fat.
P<.10.
** P <.05.


Dietary
EE,
%
1)6.7a
2)22.5a


1) 1.8
2)4.2
3)4.1


1)3.3
2)5.2
3)7.4
4)7.4
5)9.6






1)3.2
2)6.2
3)6.2
4)3.2
5)6.2


Diet, % of DM


-15-75 Pasture and 5 or 4 kg of
concentrate offered for
treatments 1 and 2,
respectively

16-112 12% corn silage, 14%
alfalfa silage, 9.5%
alfalfa hay, 64.5%
concentrate


--- 160-188 38% corn silage, 8%
alfalfa hay, 7% alfalfa
silage, 47% concentrate


50-125 4 ill ill i .h 1 57%
concentrate









21-126 30% corn silage, 20%
alfalfa silage, 50%
concentrate


DMI, Milk,
kg/d kg/d


1)20.3
2) 19.7



1)22.2
2)21.1
3)21.6


1)22.3
2) 24.0
3)22.0
2 vs. 3**


1)28.2
2) 27.2
3)28.8
4)26.8
5)24.1






1)23.6a
2) 22.7ab
3) 21.3bc
4) 21.3bc
5) 20.40


1)23.7
2) 25.0
F**


1) 34.4
2) 34.6
3)35.5


1)35.1
2)36.8
3)37.5
F*


1)32.5
2)32.6
3)35.0
4) 34.3
5)33.0
1 & 2 vs. 3
& 4**



1) 39.6
2) 40.7
3) 36.4
4)36.1
5) 39.3


4.0% FCM, Milk
kg/d protein,
%


1)22.5
2) 24.5
F**


1) 30.0
2) 29.9
3) 31.6


1)32.4
2)32.3
3)33.4
4) 34.6
5)33.0
t
1 & 2 vs. 3
& 4**


1)35.4
2)35.0
3)33.3
4) 33.9
5)35.0


1)3.12
2)3.14


1)3.1
2) 3.0
3) 3.0


1)2.97
2)2.92
3)2.86



1)3.20
2)3.03
3)3.03
4)3.08
5)3.07
1 vs. 2 *




1) 3.03a
2) 2.83b
3) 2.88bc
4) 3.08a
5) 2.98ac


Milk fat, BW change, kg
% or kg/d


1) 3.64
2) 3.86
F**


1)-0.40
2)-0.04
(kg/d)


1) 3.2
2)3.1
3) 3.3


1)3.76
2)3.83
3)3.08
F**
2 vs. 3**

1)3.49
2)3.48
3)3.26
4)3.58
5)3.51


1)0.87
2) 1.29
3)0.71
(kg/d)
2 vs. 3**

1)0.70
2)0.82
3)0.57
4) 0.64
5)0.46
(kg/d)


1) 3.33
2) 3.09
3)3.50
4) 3.63
5) 3.29


















Table 2.4. Effects of feeding ruminally inert fat (Ca salts of fatty acids (CSFA) or prilled fat (PF)) to the same cows throughout the

study on concentration of plasma hormones and metabolites.


Reference Treatments/ Fat
source

Erickson et 1) Control
al., 1992 2) 12 g/d niacin (NA)
3) 3% CSFA
4) NA + CSFA


Moallem et 1) Control
al., 2000 2) 0.55 kg/d CSFA
3) Control + bST


Garcia-
Bojalil et al.,
1998


1) 11.1% RDP
2) 11.1% RDP + 2.2%
CSFA
3) RDP 15.7%
4) 15.7% RDP + 2.2%
CSFA


Spicer et al., 1) Control
1993 2) 1.8% CSFA



Selberg et al., 1) Control
2004 2) 225 g/d CaS
conjugated linoleic
acid (CLA)
3) 225 g/d CaS trans-
C18:1


Dietary
EE,


1)4.77
2)6.65
3)4.62
4) 6.20


Control
prepartum:
4.3
Control
post-
partum: 5.2


DIM Diet, % of DM


15-98 35% alfalfa haylage,
10% corn silage,
55% concentrate



0-150 9.5% wheat silage,
15% corn silage,
3.5% legume hay,
3.5% oat hay, 2.3%
wheat straw, 66.2%
concentrate

0-120 34% corn silage,
13' i11 11 i .
53% concentrate






28-84 20% sorghum silage,
19' i1 il .
61% concentrate


-28-49 Prepartum: 13%
bermuda grass hay,
39% corn silage,
52% concentrate
Postpartum: 10%
il 11 i.. 29%
corn silage, 61%
concentrate


NEFA,
peq/L

1)265
2)238
3)303
4)352
F**


BHBA,
mg/dl

1)6.50
2)5.18
3)7.82
4) 6.48
F**


Insulin,
ng/ml


IGF-1,
ng/ml


Other


Progesterone based
cyclicity, DIM
1)27
2)29
3)26


RDP x F** a


1)0.69
2)0.60
3)0.55
4)0.52
F*


1)36.1
2) 82.7


1 vs. 2 by
week** (CaS
CLA peaked
higher at wk 1
postpartum)


1 vs. 2 by
week** (CaS
CLA peaked
higher at wk
1 postpartum)


1 vs. 3 at
week 6**
(CaS trans-
C18:1 cows
were greater)


Progesterone
1)6.03
2) 4.47
F**


Mean hepatic lipid and TAG
concentrations did not differ
among diets


aConcentrate mix included 6-15% whole cottonseed.
b NEFA concentrations from 0-49 DIM were elevated for cows fed 11.1% RDP + CSFA in comparison to 11.1% DIP, while15.7% RDP + CSFA decreased concentrations in comparison to the 15.7%
RDP group. From 0-14 DIM cows fed 15.7% RDP diet had elevated NEFA concentrations in comparison to 15.7% RDP + CSFA.
F = effect of fat.
*P <.10.
**P<.05.


















Table 2.4. Continued.
Reference Treatments/ Fat
source


Beam and
Butler,
1998


1) Control
2) 2.59% PF


Jerred et al., 1) Low silage (LS)
1990 c 2) LS + 5% PF
3) Medium silage
(MS)
4) MS + 5% PF
5) High silage (HS)
6) HS + 5% PF



Skaar et al., 1) Control
1989 2) 12 g/d niacin (NA)
3) 5% PF
4) 12 g/d NA and 5%
PF



Pickett et 1) Control
al., 2003 2) 500 ml/d propylene
glycol (PG) drench
3) 0.45 kg/d CSFA
drench
4) 500 ml/d PG + 0.45
kg/d CSFA drench


Dietary
EE,


1)4.8
2)7.0



1)3.1
2)6.5
3) 3.4
4) 7.2
5)3.8
6) 7.2


1) 3.4b
2) 3.4b
3) 11.8b
4) 11.8b


Diet, % of DM


0-100 26% corn silage,
18% alfalfa haylage,
56% concentrate


5-105 Trt 1 & 2: 45%
alfalfa silage, 55%
concentrate
Trt 3 & 4: 64%
alfalfa silage, 36%
concentrate
Trt 5 & 6; 84%
alfalfa silage, 16%
concentrate

-17-105 25% corn silage,
25% alfalfa silage,
50% concentrate


4.8 0-21 31% corn silage,
16% alfalfa hay, 9%
ill ill 44%
concentrate a
Drenches were
administered from 0-
3 DIM


NEFA,
peq/L

1)554
2)656


BHBA,
mg/dl


Insulin,
ng/ml

1)0.37
2)0.33


IGF-1,
ng/ml


Other


1)47.7
2)37.6
F*
(1-3 wk PP)


1) 13.3
2) 19.1
F**


1)298
2)343
3)320
4)371




1)643
2)503
3)602
4)558


1) 10.8
2) 11.2
3) 10.2
4)9.7




1) 10.8
2)8.1
3)9.4
4)9.4


Increase in total hepatic
lipids by fat feeding at 0
DIM and at 5 wk
postpartum (P < 0.15);
hepatic TAG = not
significant


Hepatic TAG % at 7 DIM
1) 10.8
2)6.1
3) 10.0
4) 10.0


1)0.66
2) 0.72
3)0.63
4)0.61


aConcentrate mix included 6-15% whole cottonseed.
b concentrate mix only.
c Note: there are no fat x forage interactions; results are presented as
F = effect of fat.
*P <.10.
** P <.05.


1) control (treatments 1, 3 and 5) and 2) fat (treatments 2, 4, and 6).


















Table 2.4. Continued.
Reference Treatments/ Fat
source

Schroeder et 1) TMR fed control
al., 2003 2) Pasture + 6.7 kg/d
corn based
concentrate
3) Pasture + 6.7 kg/d
concentrate with 0.8
kg CSFA

Moallem et 1) Control
al., 1999 2) 0.55 kg/d CSFA
3) Control + bST



Moallem et 1) Control
al., 1997 2) 0.5 kg/d CSFA
3) Control + bST
4) 0.5 kg/d CSFA +
bST

Sklan et al., 1) Control
1994 2) 2.5% CSFA



Cervantes et 1) Control
al., 1996 2) 0.4 kg/d CSFA
3) 12 g/d nicotinamide
(NM)
4) 0.4 kg/d CSFA + 12
g/d NM


Sklan et al., 1) Control
1991 2) 2.6% CSFA


Dietary
EE,
%
1)4.5
2)6.1
3)8.1


Diet, % of DM


117-152 Control: 59% corn
silage, 41%
concentrate
Trt 2 and 3: -84%
pasture, 16%
concentrate


--- 0-150 8% wheat silage,
20% corn silage, 3%
pea hay, 3% oat hay,
66% concentrate a

--- 0-150 10.4% wheat silage,
24% corn silage,
2.3% pea hay, 2.1%
oat hay, 61.2%
concentrate


1)2.8
2) 4.9



1)3.1
2)5.1
3)3.0
4)5.0


0-120 14.3% wheat silage,
15.3% corn silage,
6.4% vetch hay, 64%
concentrate a

112-150 Varied dependent on
stage of lactation.
Forages (35-60% of
diet) utilized were
i i ,I alfalfa
haylage, and corn
silage.


--- 0-120 13.7% corn silage,
11.3% vetch hay,
75% concentrate a


aConcentrate mix included 6-15% whole cotton seed.
F = effect of fat.
P = effect of parity.
N.S. =not significant.
*P <.10.
** P<.05.


NEFA,
peq/L

1)349
2)289
3)311


1)159
2) 152
3) 125
(measured only
at 50 DIM)
F**
(fat feeding
increased NEFA
concentrations)


1)153
2)172
FxP*


1)120
2) 157
3) 126
4) 151
F*


BHBA,
mg/dl


Insulin,
ng/ml


IGF-1,
ng/ml


Other


Progesterone, ng/ml
1)33.0
2) 55.4
3)30.0
F**
DIM to first
ovulation
1)24.5
2)26.9
3)28.4
4) 27.5
1st conception rate
was lower for
primiparous cows
fed CSFA
FxP**


1) 3.9
2) 3.9
3)3.8
4) 3.3


Fat fed had
greater
concentrations
until 40 DIM**

















Table 2.5. Effects of feeding oilseeds alone or in combination with other fat
concentration of plasma hormones and metabolites.
Reference Treatments/Fat Dietary DIM Diet, % of DM NEFA,
source EE, peq/L


Drackley et 1) Control
al., 1998 2) Control + 12 g/d
niacin
3) 10% whole raw
soybeans and 2.5%
tallow
4) Treatment 3 + 12
g/d niacin


1)2.75
2)2.75
3)6.04
4) 6.04


28-301 32.5% alfalfa
haylage, 17.5% corn
silage, 50%
concentrate a


1)98
2)117
3)134
4)122
F**


BHBA,
mg/dl

1)4.9
2)4.8
3)4.6
4)5.3
FxT*


F x niacin** F x niacin x
F x niacin x T** T**


Pires, et al., 1) Control 1)3.2 21-126 30% corn silage, 1)197
1996 2) 18% ground 2)6.2 20% alfalfa silage, 2)248
soybean 3)6.2 50% concentrate 3)200
3) 18% whole soybean 4) 3.2 4) 194
4) 2.7% blood meal 5) 6.2 5) 233
5) 2.7% blood meal +
3 % tallow
aAfter 175 DIM, diets were adjusted for decreased nutrient requirements. Forage content increased to 60% of DM. For treatments 3 and 4 whole raw soybeans were removed and tallow was decreased
to 2.25% of the diet.
F = effect of fat.
T effect of time.
*P <.10.
** P <.05.


sources to the same cows throughout the study on


Insulin,
ng/ml


IGF-1,
ng/ml


Other


















Table 2.6 Effects of feeding rendered fats to the same cows throughout the study on concentration of plasma hormones and


metabolites.
Reference Treatments/ Fat source


Bateman et al.,
1996


1) 33% NDF, 0% tallow
2) 33% NDF, 2% tallow
3) 40% NDF, 0% tallow
4) 40% NDF, 2% tallow


Bateman et al., 1) 33% NDF, 0% tallow
1996 2) 33% NDF, 2% tallow
3) 40% NDF, 0% tallow
4) 40% NDF, 2% tallow


Salado et al., 1) Control
2004 2) 0.7 kg/d partially
hydrogenated vegetable
oil
F = effect of fat.
*P <.10.
** P <.05.


Dietary
EE,
%
1)3.5
2)5.4
3)3.7
4)5.8




1)3.5
2)5.4
3)3.7
4)5.8


1)6.7 b
2) 22.5 b


DIM


Diet, % of DM


120-240, .11 .11 .1. 22% corn
winter silage, 14% alfalfa silage,
0 or 13% earlage, 56 or
43% concentrate


120-240, 11 11 1. 22% corn
summer silage, 14% alfalfa silage,
0 or 13% earlage, 56 or
43% concentrate


-15-75 Pasture and 5 or 4 kg of
concentrate offered for
treatments 1 and 2,
respectively


BHBA,
mg/dl


Insulin,
ng/ml


IGF-1,
ng/ml


Other


NEFA,
peq/L

1) 190
2) 220
3) 160
4)210
F*



1) 450
2) 510
3) 460
4) 540
F*

1) 659
2) 691


1)0.52
2)0.55


1) 140.1
2) 112.5














CHAPTER 3
EFFECTS OF THE TIMING OF INITIATION OF FEEDING CALCIUM SOAPS OF
LONG CHAIN FATTY ACIDS ON PERIPARTURIENT HOLSTEIN COWS DURING
SUMMER

Materials and Methods

Cows and Diets

Holstein cows, housed at the Dairy Research Unit near Hague, FL (290 44' N

latitude, 820 26' W longitude) were blocked by predicted calving date, parity

(primiparous or multiparous), body weight (BW), and milk production of the previous

year for multiparous cows and assigned randomly to treatment at approximately 28 d

prior to their due date. Parturition occurred between April 24 and August 9, 2003 for all

cows. A total of 58 cows were assigned to the experiment and calved having consumed

their dietary treatment for at least 10 d prior to calving. However, due to lameness (n =

3), displaced abomasum (n = 3), death due to respiratory infection (n = 1), severe mastitis

(n = 1), severe vaginal tear (n = 1), ketosis, retained fetal membranes (RFM) and static

rumen for 10 continuous d (n = 1), and an undiagnosed condition (n = 1), only 47 cows (n

= 25 multiparous and n = 22 primiparous) were included in the final data set. Sample

collections, housing conditions, and animal care met the requirements of the "Animals

Used for Teaching and Research Protocol" approved by the Animal Care and Use

Committee at the University of Florida, Gainesville.

Animals were housed on pasture with shade and cooling fans provided beginning at

approximately 28 d prior to parturition. At approximately 7 d prior to parturition, cows

were moved to a sand-bedded barn with cooling fans and sprinklers to allow for close









observation of signs of impending parturition. After parturition cows were housed in an

open-sided free stall barn equipped with sand bedding, cooling fans, sprinklers and self-

locking stanchions where they were fed their assigned dietary treatment.

Two diets were prepared and fed as a total mixed ration (TMR) (Table 3.1 and 3.2)

in ad libitum amounts twice daily to allow for 5 to 10% refusal in the prepartum and

postpartum periods, respectively. Calcium salts of long chain fatty acids (CSLCFA,

Megalac-R, Church & Dwight Co., Princeton, NJ) were fed at 0 or 2.0% of dietary dry

matter (DM). The fatty acid (FA) profile of CSLCFA provided by the manufacturer was

17.4% C16:0, 2.1% C18:0, 32.1% C18:1 cis, 1.5% C18:1 trans, 30.5% C18:2, 2.4%

C18:3, and 12.2% other FA. Four experimental treatments were the following: 0%

CSLCFA (Control), CSLCFA fed starting at 28 d prepartum, CSLCFA fed starting at 1 d

in milk (DIM), and CSLCFA fed starting at 28 DIM. Once initiated, all diets continued

through 100 DIM. Those cows receiving CSLCFA beginning in the postpartum period

were fed CSLCFA at 1% of dietary DM for 7 d in order to adapt cows to CSLCFA

slowly.

Sample Collection

Representative samples were obtained of corn silage, bermudagrass hay, alfalfa

hay, and concentrate mixes on a weekly basis. Corn silage was immediately dried at

55C for 48 h in a forced air oven in order to calculate concentration of DM and maintain

the same formulated forage to concentrate ratio. The weekly samples were composite

on a monthly basis, ground through a 1-mm Wiley mill screen (A. H. Thomas,

Philadelphia, PA) and analyzed for chemical composition using wet chemistry (Dairy

One, Ithaca, NY). Cows were milked three times per day at 0200, 1000, and 1800 h.

Milk production was recorded at each milking. Milk samples were collected weekly









from two consecutive milkings and analyzed by Southeast Milk lab (Belleview, FL) for

fat, true protein, and somatic cell count (SCC) using a Bently 2000 NIR analyzer. Body

weight was measured weekly after the 0900 h milking. Body condition scores (BCS)

were assigned by the same two individuals at -4, 0, 3, 6, 9, 12, and 14 wk postpartum

(Edmonson et al., 1989). Blood was collected three times weekly from parturition until

artificial insemination (AI) (72 3 DIM) and again at 7 d post AI from the coccygeal or

the jugular vessels immediately before the 1000 h milking using 13 x 100 ml vacutainer

tubes containing sodium heparin (Becton Dickinson Vacutainer systems, Franklin Lakes,

NJ). Samples were put immediately on ice until centrifuged at 2619 g at 50C for 30 min

(RC-3B refrigerated centrifuge, H 600A rotor, Sorvall Instruments, Wilmington, DE).

Plasma was decanted and frozen at -200C. Liver samples were collected at 2, 14 1, and

28 1 DIM via liver biopsy, rinsed with sterile saline and immediately frozen in liquid

nitrogen (-1920C) and stored at -800C.

Reproductive Management

The previous pregnant uterine horn was determined as the longer horn with the

greater diameter using a real time Ultrasound Aloka 500 scanner (Aloka Co., Ltd, Tokyo,

Japan) equipped with a 5.0 MHz linear rectal transducer. The diameters of the uterine

horn and cervix were measured at 21 + 3 and 28 3 DIM. The transducer was placed in

a transverse position in relation to the horns, at approximately 4 cm past the bifurcation

of the horns. When the transducer was positioned and the horns could be seen clearly,

the image was fixed. Pressure with the transducer on the uterine horns was avoided in

order to obtain a circular cross-section image of the horns. Machine calipers were

activated such that a vertical line was extended from serosa to serosa of the uterine horn

cross-section. Cervical diameter was measured by placing the transducer in a transversal









position in relation to the cervix at its middle section and the distance between two points

was obtained as described above. A 7.5 MHz linear rectal transducer was used to

monitor ovarian structures. Size, location and number of follicles were recorded at 21

and 28 DIM and categorized by size into Class 1 (2 to 5 mm), Class 2 (6 to 9 mm), and

Class 3 (> 10 mm) follicles (Lucy et al., 1992). The location and size of corpus luteum

(CL) (determined by measurement of length and width) were recorded.

At 21 3 and 28 3 DIM, uterine tonus was classified as none, moderate, and

intense. Also, vaginal examinations were conducted to characterize uterine condition at

21 3 and 28 3 DIM using a glass speculum inserted into the vagina until the cervical

os could be seen. The cervix was classified as open or closed, the cranial vagina was

classified as pink or red, and vaginal discharge was classified according to amount (none,

trace, slight, moderate, or copious) and quality (none, clear mucous, cloudy mucous,

mucopurulent, or purulent).

Cows were enrolled in a Pre-synchronization/Ovsynch protocol beginning at 44 +

3 DIM by injecting gonadatropin releasing hormone (GnRH) (100 atg, Gonadorelin

Diacetate Tetrahydrate, Cystorelin, Merial Ltd., Athens, GA) and ovaries were scanned

every other day for 7 d. At 51 3 DIM, prostaglandin F2a (PGF2,) (25 mg, Lutalyse,

Pharmacia Upjohn, Kalamazoo, MI) was injected. Ultrasound examination of the ovaries

continued until the formation of a new CL. Eleven days later (62 + 3 DIM), GnRH was

injected again (100 atg), followed by PGF2a (25 mg) 7 d later (69 + 3 DIM), and GnRH

(100 [tg) 48 h following PGF2, (71 3 DIM). Ovaries were scanned by ultrasonagraphy

at each injection day. All hormone injections were given intramuscularly (i.m.) after a

blood collection from coccygeal vessels. Cows were inseminated at 72 3 DIM (16 h









following GnRH) using the same batch of semen by the same trained inseminator.

Immediately after AI, an injection of bST (POSILAC Monsanto Co., St. Louis, MO)

was given in the ischiorectal fossa and continued every 14 d thereafter. At 7 d post AI

(79 3 DIM), ovaries were examined by ultrasound for the presence of a CL. Cows with

no CL were reenrolled in Ovsynch. At 28 d post AI (100 3 DIM), pregnancy was

determined by identifying the presence of embryonic fluid, appearance of embryo and an

embryonic heart beat using ultrasound. Pregnancy was confirmed by rectal palpation at

45 and 72 d post AI by the farm veterinarian.

Animal Health

Body temperature was measured the first 5 d postpartum using a rectal

thermometer. Cows were diagnosed as having RFM if membranes were still attached 24

h after parturition. Treatment of RFM was penicillin (50 cc, i.m.) for 5 d. Cows having a

displaced abomasum had corrective surgery performed by the farm veterinarian, followed

by administration with dextrose (500 ml) and vitamin B complex (15 cc) intravenously

plus calcium propionate (300 ml) and 30 g of Probios (Vets Plus Inc., Knapp, WI) orally.

Ketosis was diagnosed using Ketostix (Bayer, Pittsburgh, PA). Treatment for ketosis

was dextrose (500 ml) and vitamin B complex (15 cc) intravenously and 30 g of Probios

(Vets Plus Inc., Knapp, WI) orally. Cows having a rectal temperature > 39.40C and with

no other apparent health conditions were diagnosed as having metritis and treated with

0.23 to 0.45 mg/kg BW ofNexcel (ceftiofur sodium sterile powder, Pfizer Inc., New

York, NY).

Sample Analysis

Plasma concentrations of nonesterified fatty acids (NEFA) (NEFA-C kit; Wako

Fine Chemical Industries USA, Inc., Dallas, TX; as modified by (Johnson, 1993) and P-









hydroxy butyric acid (BHBA) (Pointe Scientific Inc., Lincoln Park, MI) were determined

once weekly. A Technicon Autoanalyzer (Technicon Instruments Corp., Chauncey, NY)

was used to determine weekly concentrations of blood urea nitrogen (BUN) (a

modification of (Coulombe and Favreau, 1963) as described in Bran + Luebbe Industrial

Method #339-01) and plasma glucose (a modification of(Gochman and Schmitz, 1972)

as described in Bran + Luebbe Industrial Method #339-19).

A double antibody radioimmunoassay (RIA) was used to determine plasma

concentrations of insulin (Badinga et al., 1991; Malven et al., 1987) and IGF-1 (Badinga

et al., 1991) on every plasma sample collected. The sensitivity of the insulin assay was

0.3 ng/ml, and intra- and interassay coefficients of variation (CV) were 9.1 and 14.9%,

respectively. The sensitivity of the IGF-1 assay was 50 pg/ml, and intra- and interassay

CV were 11.4 and 12.1%, respectively. Weekly leptin concentrations were determined

by RIA at the University of Missouri (Delavaud et al., 2000). Intra- and interassay CV

were less than 10%. Concentrations of plasma progesterone were determined on every

plasma sample collected using Coat-A-Count Kit (DPC Diagnostic Products Inc., Los

Angeles, CA) solid phase 1251 RIA. The sensitivity of the assay was 0.1 ng/ml and the

intra- and interassay CV were 0.8 and 5.2%, respectively. A polyethylene glycol RIA

procedure described by Meyer et al. (1995) was used to analyze the concentration of 15-

keto-13,14-dihydro-prostaglandin F2 (PGFM) on daily plasma samples collected during

the first 10 DIM. The sensitivity of the assay was 31.25 pg/ml and the intra- and

interassay CV were 8.2 and 20.6%, respectively.

The three plasma samples collected weekly were composite into one sample

during each of the first 4 wk postpartum and analyzed for concentrations of alkaline









phosphatase (ALK) (Diagnostic Chemicals Ltd, Oxford, CN), alanine aminotransferase

(ALT) (Abbott Diagnostics, Abbott Park, IL), aspartate aminotransferase (AST) (Abbott

Diagnostics, Abbott Park, IL), gamma glutamyl transferase (GGT) (Thermo DMA,

Arlington, TX), albumin (Diagnostic Chemicals Ltd, Oxford, CN), and total biliruben

(Wako Chemicals USA, Inc., Richmond, VA) using a Hitachi 911 chemistry analyzer

(Roche Diagnostics, Indianapolis, IN).

Acute phase protein concentrations were determined on two samples per week for 4

wk postpartum. Plasma fibrinogen was determined from a standard curve generated

using a human reference (Sigma Diagnostics). Plasma haptoglobin concentrations were

determined by measuring haptoglobin/hemoglobin completing (Makimura and Suzuki,

1982). Ceruloplasmin oxidase activity was measured using colorimetric procedures

described by Demetriou et al. (1974).

Total liver cellular RNA was isolated from samples collected at 2, 14, and 28 DIM

from eight cows in the control, fat pre, and fat 1 DIM groups (n = 72) using TRIzol

reagent (Life Technologies, Grand Island, NY) according to the manufacturer's

directions. Ten [tg of RNA was fractionated in a 1.0% agrose-formaldehyde gel and

transferred overnight to BioTrans nylon membranes (ICN, Irvine, CA) by capillary

action. The RNA was cross linked to the membrane by exposure to ultraviolet light for

90 sec and the membrane was heated at 800C for 1 h. The membranes were

prehybridized for 30 min with buffer (ULTRAhyb buffer, Ambion Inc., Austin, TX) to

block non-specific binding sites. Membranes were then hybridized to 32P-labeled IGF-I,

IGF-II, and IGFBP-2 cDNA probes, respectively. After hybridization, filters were

washed for 15 min in 50 ml of 2X SSC, 0.1% SDS at 500C, followed by two 15-min









washes in 50 ml of0.1X SSC, 0.1% SDS at 500C. Filters were blotted dry and exposed

to x-ray film (Super RX, Fugi Film, Japan) for 3 to 96 h at -800C. Hybridization signals

were quantified by densitometric analysis (Eastman Kodak, Rochester, NY).

Triacylglycerol (TAG) concentrations in liver samples (75 to 150 mg, wet weight)

were determined colorimetrically (Foster and Dunn, 1973) by first extracting the total

lipid as described by Drackley et al. (1992). Dry matter content of liver (-100 mg) was

determined by drying in a forced air oven for 48 h at 550C.

Statistical Analysis

Data were analyzed as a completely randomized design using the PROC MIXED

procedure for repeated measurement of SAS (SAS software statistics, 2001) according to

the following mathematical model:

Yijkl = p + Di + Pj + DPij +Ck (ij) +W1 + DWil + PWji + DPWiikl + Eijkl

where Yijkl is the observation, t is the overall mean, Di is the fixed effect of diet (i

= 1, 2, 3, and 4), Pj is the fixed effect of parity (j = 1 and 2), DPij is the interaction of diet

and parity, Ck(ij) is random effect of cow within diet and parity (k = 1, 2, ...n), Wi is the

fixed effect of week (1= 0, 1, 2, ...14), DWil is the interaction of diet and week, PWji is

the interaction of parity and week, DPWij1 is the three way interaction of diet, parity and

week, and Eijkl is the residual error.

Results are reported as least square means. Significance was determined at P <

0.05 and tendencies included P > 0.05 and < 0.10. Orthogonal contrasts used included 1)

no CSLCFA versus CSLCFA (control vs. (CSLCFA pre, CSLCFA 1 DIM, plus CSLCFA

28 DIM)), 2) CSLCFA feeding initiated prepartum vs. CSLCFA feeding initiated

postpartum (CSLCFA pre vs. (CSLCFA 1 DIM plus CSLCFA 28 DIM)), and 3)

CSLCFA initiated at 1 DIM versus CSLCFA initiated at 28 DIM. These three contrasts









by parity interactions were tested also. In addition, for dependent variables measured

only before 28 DIM, the cows assigned to the CSLCFA 28 DIM treatment were

combined with control cows. For this data set (when only three treatments had been

initiated), a separate set of orthogonal contrasts was used. Contrasts were 1) CSLCFA

feeding initiated in the prepartum vs. no CSLCFA feeding in the prepartum period

(CSLCFA pre vs. (Control plus CSLCFA 1 DIM plus CSLCFA 28 DIM)), and 2) no

CSLCFA (Control plus CSLCFA at 28 DIM) vs. CSLCFA initiated at 1 DIM. These two

contrasts by parity interactions were tested also.

Data for NEFA, BHBA, BUN, glucose, leptin, alkaline phosphatase, alanine

aminotransferase, aspartate aminotransferase, gamma glutamyl transferase, albumin, total

biliruben, hepatic mRNA, hepatic TAG, milk production, BW, and BCS were evaluated

using repeated measures of the Mixed procedure of the SAS software program. Data were

tested to determine the structure of best fit, namely AR (1), ARH (1), CS, or CSH, as

indicated by a lower Schwartz Baesian information criterion value (Littell et al., 1996).

Orthogonal contrasts (cited above) were used to determine if the mean values were

different. If main effects or contrasts were significant, the slice command was used to

determine at which time points the treatments differed. If slice was significant, then the

pdiff command was used to determine which treatments were different at that time point.

Data that did not have a common day in milk (insulin, IGF-I, P4, PGFM,

fibrinogen, haptoglobin, and ceruloplasmin) were modeled using the CS structure as a

polynomial function of time using regression analysis and coefficients were obtained to

plot the curves after the level of order (linear, quadratic, cubic, quartic, or quintic) that









best fit the data was determined. Heterogeneity of regression was performed to

determine if the curves differed for each of the orthogonal contrasts cited above.

Data for average milk protein concentration, average milk fat concentration,

average milk protein yield, average milk fat yield, 3.5% fat corrected milk (FCM) yield,

vaginal exams, and 21 DIM and 28 DIM ovarian maps were analyzed using the Proc

GLM procedure of SAS.

Odds ratios and their confidence intervals were obtained using the Chi square and

PROC LOGISTIC procedures for testing of first service conception rates, estrus cycling

rates (the first day postpartum of ovulation was determined by plasma progesterone

concentrations > 1 ng/ml for two consecutive sampling days) and incidence of disease.

Results and Discussion

Dry Matter Intake and Diets

Concentrations of nutrients in experimental diets were within or exceeded the

acceptable targets for close-up nonlactating cows and lactating cows at 90 DIM (NRC,

2001) (Tables 3.1 and 3.2). The ether extract values of the diets containing CSLCFA

increased 1.3 and 1.6 percentage units for the close-up and lactating diets respectively.

During the prepartum period, cows consumed an average of 10.0 and 10.2 kg of

DM/d when offered the 0 or 2% CSLCFA diets, respectively. Cows were fed diets an

average of 24.8 + 7.4 d prior to parturition. During the postpartum period, cows

consumed an average of 17.5 and 16.4 kg of DM/d when offered the 0 or 2% CSLCFA

diets, respectively. In a review of several studies, Allen (2000) reported a linear decline

in DMI of 2.5% for every 1% inclusion of CSLCFA in the diet DM. Although not

statistically evaluated, the difference of 6.3% is close to the 5% depression of intake

predicted by Allen's (2000) equations.









Production and Body Weight

As expected, multiparous cows were heavier (644 vs. 530 kg; P = 0.001) and

produced more milk (43.4 vs. 41.0 kg/d; P = 0.041) than primiparous cows (Table 3.3).

In addition, multiparous cows tended to have a greater SCC than did primiparous cows

(586 vs. 354 x 1000/ml; P = 0.067; Table 3.3), however no treatment differences were

detected.

It is well documented that milk production often increases when supplemental fats

are fed. This may be due to increased energy intake. In the current study, cows fed

CSLCFA in the prepartum period tended to produce more milk (42.2 vs. 37.1 kg/d; P =

0.059) and 3.5% FCM (41.3 vs. 36.5 kg/d; P = 0.087) the first 100 DIM than cows fed

CSLCFA beginning in the postpartum period (Table 3.3). Feeding CSLCFA beginning

prepartum probably increased milk production due to increased energy intake and

because the cows did not have the additional stress of adjusting to a new dietary

ingredient early in lactation. Frequently there is a three to four wk delay before milk

production increases when supplemental fats are added during the postpartum period

(Garcia-Bojalil et al., 1998; Schingoethe and Casper, 1991). In contrast to the results of

this study, Salfer and coworkers (1995) did not detect any difference in milk yield or

3.5% FCM yield of cows fed diets of 0 or 2% partially hydrogenated tallow (PHT)

beginning at 14 d prepartum, at 1 DIM, or at 35 DIM through 151 DIM. However, when

only the first 35 DIM were considered, yield of 3.5% FCM by cows fed PHT beginning

at 1 DIM tended to be greater than that of cows fed PHT beginning 14 d prepartum (33.4

vs. 36.9 kg/d).

Milk protein concentration was reported to not change (Atwal et al., 1990; Markus

et al., 1996; Moallem et al., 2000) or to decline when supplemental fats were fed









(AbuGhazaleh et al., 2004; Cervantes et al., 1996; Drackley et al., 1998). Protein

concentration in the milk of primiparous cows was 2.63% for both no CSLCFA and

CSLCFA fed groups, however multiparous cows fed CSLCFA had increased milk protein

concentration in comparison to cows not fed CSLCFA (2.70 vs. 2.47%) (no CSLCFA vs.

CSLCFA by parity interaction, P = 0.091). This increase could be due to more ruminal

microbes delivering protein for protein synthesis by the mammary gland. When fat is

fed, bacteria numbers may increase in concurrence with a decrease in protozoal

populations (Sutton et al., 1983). Likely due to increased milk production, milk protein

yield tended to be greater by cows fed CSLCFA prepartum in comparison to cows fed

CSLCFA beginning postpartum (1.15 vs. 1.02 kg/d; P = 0.08).

In this study, no treatment or parity differences were detected for average milk fat

concentration (3.44%) or yield (1.32 kg/d) through 100 DIM. In contrast, some studies

reported an increase in milk fat concentration when CSLCFA were included in the diet

(Cervantes et al., 1996; Moallem et al., 2000; Sklan et al., 1994; Sklan et al., 1991). The

response of milk fat concentration to dietary fat is dependent upon many factors

including the fat concentration and composition in the basal diet and in the supplement as

well as the forage source and amount. The ruminally inert fat fed in this study should not

have interfered with ruminal digestion and the diet included sufficient fiber from both

corn silage and alfalfa hay. About 50% of the fat found in milk is synthesized in the

mammary gland from acetate and butyrate, while the other 50% comes directly from fat

absorbed from the blood (Ackers, 2002). Increased uptake of long chain FA by the

mammary gland for milk fat synthesis may inhibit the synthesis of short and medium

chain FA (Bauman and Griinari, 2003). Because we did not analyze the FA present in









the milk, we can only speculate that there was a balance between increased dietary FA

uptake and decreased synthesis of short and medium chain FA in the mammary gland

such that total concentration of milk fat did not differ from control cows. Palmquist and

coworkers (1993) published an equation based on 49 published experiments predicting an

increase in milk fat concentration of 0.18% due to feeding an additional 0.5 kg of fat

daily. In the present study, milk fat concentration was 0.11% units greater for cows fed

CSLCFA versus controls however this was not significantly different.

In agreement with several other studies in which fat supplementation was initiated

in the periparturient period (Garcia-Bojalil et al., 1998; Kim et al., 1993; Moallem et al.,

2000), treatment did not affect mean postpartum BW (587 kg). Multiparous cows not fed

CSLCFA continued to gradually lose BW for 12 wk whereas those fed CSLCFA reached

nadir by two to three wk postpartum. The pattern of BW for primiparous cows appeared

similar for all treatment groups (treatment by parity by week interaction, P < 0.001,

Figure 3.2). A treatment by week interaction was detected for body condition score (P =

0.007, Figure 3.3). Cows fed CSLCFA beginning at 1 or 28 DIM reached their nadir

sooner and began gaining body condition earlier postpartum than controls or cows fed

CSLCFA beginning prepartum. Greater loss of body condition by these latter two groups

of cows was most likely due to less energy intake and greater milk yield respectively.

Plasma Metabolites

Typically, plasma BHBA concentrations are not affected by fat supplementation,

but can decrease slightly in fat-supplemented cows if cows on the basal diet have high

BHBA concentrations (Grummer and Carroll, 1991). In this study, mean plasma

concentration of BHBA was lower for cows fed CSLCFA in comparison to controls (8.7

vs. 12.5 mg/dl, respectively; P = 0.007; Table 3.4). This difference was particularly









apparent for control versus CSLCFA 1 DIM and 28 DIM at wk 3, 4, and 5 postpartum

and at wk 5 for CSLCFA prepartum (P < 0.05; Figure 3.4). Control cows may have been

mobilizing more of their adipose tissue early in lactation to support milk production due

to the lower energy dense diet than cows fed CSLCFA. In addition, an interaction of

treatment and parity was detected (Table 3.4). Multiparous cows fed CSLCFA had lower

concentration of plasma BHBA than control cows whereas that of primiparous cows was

greater but unaffected by diet (no CSLCFA vs. CSLCFA by parity interaction, P =

0.044).

When fat is supplemented, plasma concentrations ofNEFA routinely increase

(Drackley, 1999), although there are several reports of numeric increases in NEFA

concentrations that are not significant (Beam and Butler, 1998; Salado et al., 2004). In

this experiment, control cows did not differ from cows fed CSLCFA, probably due to

control cows and CSLCFA prepartum following a similar pattern while cows fed

CSLCFA beginning at 1 DIM and 28 DIM were similar but followed a different pattern.

Mean plasma concentrations of NEFA were greater for cows fed CSLCFA in the

prepartum period in comparison to cows fed CSLCFA beginning postpartum (456 vs. 294

meq/L; P = 0.002). This difference was particularly evident at wk 3 and 4 postpartum (P

< 0.05, Figure 3.5). A large portion of the plasma NEFA in cows fed fat prepartum may

have been from adipose mobilization to support greater milk production because plasma

BHBA followed a similar pattern although not significant. Grum et al. (1996) reported

lower concentrations of plasma NEFA postpartum when cows were fed supplemental fat

(Qual-Fat) beginning prepartum in comparison to cows not fed supplemental fat

prepartum.









Concentrations of plasma glucose are not affected generally by fat supplementation

(Grummer and Carroll, 1991). However in this study, concentration of plasma glucose

was increased in cows fed CSLCFA in comparison to controls (65.6 vs. 60.4 mg/dl; P =

0.004) but this was observed mainly in multiparous cows. Similar to what was observed

with BHBA, multiparous cows not fed CSLCFA tended to have lower concentrations of

plasma glucose than those fed CSLCFA (55.2 vs. 65.5 mg/dl) but primiparous cows

across diets did not differ in plasma glucose (no CSLCFA vs. CSLCFA by parity

interaction, P = 0.054). Elevated blood ketones are associated often with lowered blood

glucose. In addition, concentration of plasma glucose was lower for cows fed CSLCFA

beginning prepartum versus postpartum (62.8 vs. 67.0 mg/dl; P = 0.025) and was greater

for primiparous cows versus multiparous cows (66.8 vs. 61.7 mg/dl; P = 0.001). A

tendency for an interaction of treatment and week was detected (P = 0.10) and treatment

differences were evident at wk 3, 4, 5, and 6 postpartum (P < 0.05; Figure 3.6). Control

cows had lower concentrations of plasma glucose than those fed CSLCFA at 1 DIM and

28 DIM at wk 3, 4, and 5 postpartum, than those fed CSLCFA prepartum at wk 5

postpartum, and than those fed CSLCFA at 1 DIM at wk 6 postpartum. Cows fed

CSLCFA prepartum had lower plasma concentrations of glucose than those fed CSLCFA

at 28 DIM at wk 3 and lower than those fed CSLCFA at 1 DIM at wk 6 postpartum.

Differences in plasma glucose concentrations may be due to differences in DMI and milk

production between treatments and parities. In a review of dietary fat and adipose tissue

metabolism, Chilliard (1993) noted that in 52 comparisons the difference in plasma

glucose between control and fat-supplemented groups was 0 g/L (+ 0.03).









Mean plasma concentrations of BUN tended to be greater for cows fed CSLCFA

starting prepartum in comparison to cows fed CSLCFA starting postpartum (12.9 vs. 10.6

mg/dl; P = 0.053). This was especially true for multiparous cows (15.3 vs. 10.9 mg/dl)

compared to primiparous cows (10.4 vs. 10.2 mg/dl) (CSLCFA prepartum vs. CSLCFA

postpartum by parity interaction, P = 0.081). A tendency for a treatment by week

interaction was detected (P = 0.06; Figure 3.7), however multiple means contrasts did not

detect any differences among means. Individual DMI were not measured however milk

production was greater by cows fed CSLCFA beginning prepartum, so we can speculate

that cows fed CSLCFA beginning prepartum were consuming more DM and therefore

more dietary protein than cows fed CSLCFA beginning postpartum.

Plasma Hormones

Leptin is a hormone synthesized by adipose tissue that is stimulated by adiposity

and inhibited by undernutrition. Concentrations of leptin were decreased around the time

of parturition in concurrence with negative energy balance and a reduction in adipose

stores and may have been mediated by the reduction in plasma insulin (Block et al.,

2003). Primiparous cows fed CSLCFA tended to have a lower concentration of plasma

leptin in comparison to controls (2.15 vs. 3.06 ng/ml) whereas multiparous cows fed

CSLCFA tended to have greater concentrations of leptin in comparison to controls (2.39

vs. 1.65 ng/ml; Table 3.4; Figure 3.8) (no CSLCFA vs. CSLCFA by parity interaction, P

= 0.078). Low concentrations of circulating plasma leptin were correlated highly with

greater milk production (Liefers et al., 2003). This relationship appears to fit with

animals not fed CSLCFA; that is the higher producing multiparous cows had lower

plasma concentrations of leptin compared to the lower producing primiparous cows (1.65

vs. 3.06 ng/ml). However when CSLCFA were included in the diet, this inverse









relationship disappeared. Increasing the triglyceride concentration of the blood may

possibly influence leptin synthesis and release.

Insulin-like growth factors play a diverse role physiologically and are very

dependent upon the nutritional state of the animal. Plasma IGF-I concentrations are

correlated positively with body condition and DMI. During the periparturient period

when cows are often in a negative energy balance, circulating concentrations of IGF-I are

low (Vega et al., 1991). As the cow continues through lactation and a positive energy

status is restored, circulating concentrations of IGF-I will increase. Low IGF-I

concentrations were associated with an extended postpartum interval to estrus in beef

cows and also with delayed puberty (Roberts et al., 1997; Rutter et al., 1989), indicating

that IGF-I can be correlated positively with reproductive performance. Insulin-like

growth factor I acts synergistically with luteinizing hormone (LH) to promote follicular

development (Lucy, 2001). Beam and Butler (1998) reported lower mean concentrations

of plasma IGF-I (37.6 vs. 47.7 ng/ml) from wk 1 to 3 postpartum in lactating dairy cows

fed a diet of 2.6% prilled fat compared to no supplemental fat despite no differences in

energy balance. However, other studies reported no differences in concentration of

plasma IGF-I when supplemental fat was fed (Salado et al., 2004; Spicer et al., 1993).

In the present study, primiparous cows had a greater mean concentration of IGF-I

in comparison to multiparous cows when consuming the control diet (77.1 vs. 54.3

ng/ml); however when CSLCFA were fed, primiparous cows had a lower mean

concentration (77.1 vs. 64.2 ng/ml) whereas concentration of IGF-I increased in

multiparous cows (54.3 vs. 68.2 ng/ml) (interaction of treatment and parity; P = 0.040)

(Table 3.4). Concentrations of IGF-I were lowest immediately postpartum for all









treatments and rose throughout the lactation period. The quadratic pattern of plasma

IGF-I concentration over time was different for animals fed the control diet and animals

fed CSLCFA (P < 0.01; Table A-i) and for animals fed CSLCFA beginning prepartum

versus beginning postpartum (P < 0.01; Table A-i). Likewise, the pattern over time was

different for cows fed the control diet and cows fed CSLCFA by parity (P < 0.05; Table

A-4) and for cows fed CSLCFA beginning prepartum versus beginning postpartum by

parity (P < 0.01; Table A-4). Primiparous cows fed no CSLCFA experienced a more

rapid increase in concentrations of plasma IGF-I after parturition than CSLCFA-fed cows

before reaching a plateau at 49 DIM; however plasma IGF-1 of cows fed CSLCFA

continued to rise over time (Figure 3.9A). At 77 DIM there was no difference in

concentrations of plasma IGF-I between the two treatment groups. Multiparous cows fed

CSLCFA had greater plasma IGF-I concentrations at parturition than cows not fed

CSLCFA and their concentrations rose steadily over time so that at 77 DIM, cows fed

CSLCFA had a much greater concentration than control cows (Figure 3.9B).

Concentrations of plasma IGF-I of primiparous cows fed CSLCFA beginning prepartum

versus postpartum differed little over time (Figure 3.10A). However, multiparous cows

fed CSLCFA beginning postpartum had greater concentrations of IGF-I at parturition and

rose slightly but steadily over time whereas those fed CSLCFA beginning prepartum rose

at a greater rate starting at 42 DIM to surpass cows fed CSLCFA beginning postpartum at

77 DIM (Figure 3.10B). The differences in circulating IGF-I concentrations may have

been due to differences in energy balance.

Fat supplementation has had mixed results on circulating concentration of plasma

insulin (Staples et al., 1998). Florida workers (Garcia-Bojalil et al., 1998) reported









decreased insulin concentrations when periparturient cows were supplemented with

CSLCFA at 2.2% of dietary DM, however others reported no difference between fat-fed

cows and controls (Beam and Butler, 1998; Salado et al., 2004). Insulin has also

stimulated ovarian follicle cell growth. When granulosa cells from small (1 to 5 mm)

follicles were cultured, the addition of insulin increased cell numbers several fold and

increased progesterone production in comparison to the control (Langhout et al., 1991).

In the present experiment, plasma insulin concentrations increased gradually as week

postpartum increased (Figure 3.11), however no treatment or parity effects for mean

plasma concentration of insulin were detected (0.65 ng/ml, Table 3.4). Over time, eight

individual quadratic curves for each treatment by parity combination fit the data

significantly better than one pooled curve (Figure 3.11). However the orthogonal

contrasts for treatment, parity, and treatment by parity were not significant for plasma

concentrations of insulin over time (Table A-4). Although there were no treatment

differences, it is noteworthy that plasma insulin and glucose concentrations followed a

similar pattern.

Dietary fats typically increase concentrations of circulating cholesterol, the

precursor of progesterone (Grummer and Carroll, 1991). Ruminants fed supplemental fat

often have a slight increase in blood progesterone concentration (Staples et al., 1998).

Progesterone, secreted by the CL, prepares the uterus for implantation of the embryo and

helps maintain pregnancy by providing a nourishing environment for the concepts. At

breeding, greater concentrations of plasma progesterone has been associated with higher

conception rates (Butler et al., 1996). Work by Hawkins et al. (1995) suggested that the

increase seen in circulating progesterone when cows were fed supplemental fat was from









a reduced rate of clearance of progesterone rather than from an increased synthesis of

progesterone. Son et al. (1996) reported greater blood cholesterol and peak plasma

progesterone concentrations during the second ovulatory cycle in cows fed tallow at 2 vs.

0% of dietary DM accompanied by a tendency of improved conception. Workers at the

University of Florida (Garcia-Bojalil et al., 1998) reported that accumulated plasma

progesterone from 0 to 50 DIM was greater, pregnancy rates improved, and energy status

did not change when cows were fed diets of 2.2% calcium salts of palm oil compared to

non fat-supplemented cows.

The cubic pattern over time of plasma accumulated progesterone was different

between parities when cows were fed the control versus the CSLCFA-supplemented diet

(P < 0.05; Table A-4). Primiparous cows fed CSLCFA had a slightly greater rise in

plasma concentration of accumulated progesterone beginning at 25 DIM compared to

cows fed no CSLCFA whereas multiparous cows fed no CSLCFA had a slightly greater

rise in plasma concentration of accumulated progesterone beginning at 23 DIM than cows

fed CSLCFA (Figure 3.12). Cows were inseminated at 72 + 3 DIM when there was very

little difference in accumulated progesterone concentrations among treatment and parity

groups. Patterns over time were detected to be different between parities when fed

CSLCFA beginning at 1 DIM or at 28 DIM (P < 0.01; Table A-4). Upon closer

examination of the data, two primiparous cows fed CSLCFA beginning at 1 DIM did not

ovulate until 40 and 60 DIM (determined by two consecutive plasma progesterone

concentrations > 1.0 ng/ml) which could have skewed the accumulated progesterone

concentrations of the group. To determine if differences existed before cows were

enrolled in a synchronization program, accumulated plasma progesterone concentrations









before the first hormone injection (46 DIM 3) were analyzed. The proportion of cows

that ovulated before the first GnRH injection (44 3 DIM) were 91% (10/11), 75%

(9/12), 83% (10/12) and 100% (12/12) for control, CSLCFA prepartum, CSLCFA at 1

DIM, and CSLCFA at 28 DIM respectively and did not differ. The DIM at first ovulation

(Table 3.4) did not differ among treatments either (mean of 27 DIM). The quadratic

pattern over time differed for accumulated plasma progesterone concentrations of cows

fed the control diet and cows fed CSLCFA by parity when only 1 to 46 DIM were

included (P < 0.01; Table A-4; Figure 3.13); however patterns over time were not

different between cows fed CSLCFA beginning at 1 DIM and at 28 DIM (Table A-4).

We concluded that the difference in accumulated progesterone over time between cows

fed CSLCFA beginning at 1 DIM and at 28 DIM was not different but was due to two

animals that experienced delayed ovulation.

Through a series of desaturases and elongases, linoleic acid (LA) (C18:2) can form

dihomo-k-linolenic acid, a direct precursor to the series 1 prostaglandins, or can be

further desaturated to arachidonic acid (C20:4), a direct precursor to the 2 series

prostaglandins (Biochemistry of Lipids, Lipoproteins, and Membranes, 1996).

Prostaglandin F2,, synthesized by endometrial tissue, is an important regulator of

parturition and the estrous cycle by causing regression of the CL. Immediately

prepartum, PGF2J is important in regressing the CL of pregnancy and circulating PGF2,

concentrations decline as the postpartum uterus declines in size. Concentrations of

PGFM may be associated with immune functions such as cellular immunity and

neutrophil function. If LA is supplemented in the diet prepartum, more arachidonic acid









may be synthesized leading to higher concentrations of the series 2 prostaglandins and

possibly a greater immune competence.

Using heterogeneity of regression, the cubic patterns of plasma concentrations of

PGFM over time were different for cows fed CSLCFA prepartum in comparison to cows

not fed CSLCFA prepartum (P < 0.05, Table A-2). However parity status also influenced

this contrast (parity by no CSLCFA prepartum vs. CSLCFA prepartum interaction, P <

0.05, Table A-5). Primiparous cows fed CSLCFA prepartum had similar initial plasma

PGFM concentrations as primiparous cows not fed CSLCFA prepartum, however peak

concentrations (at -4 DIM) were greater for cows fed CSLCFA prepartum and declined

at a slower rate (Figure 3.14A). Multiparous cows not fed CSLCFA prepartum had high

initial plasma PGFM concentrations, continued to decline and stabilized at 10 DIM.

Multiparous cows fed CSLCFA prepartum had peak concentrations of plasma PGFM at 4

DIM, slowly declined and stabilized at 12 DIM (Figure 3.14B). The reason why there

was an interaction of treatment and parity is unclear. When cows that experienced RFM

and/or metritis were excluded from this analysis, the patterns and statistical significance

were unchanged. Synthesis of PGFM in cows fed diets high in LA prepartum was greater

postpartum possibly due to the increased intake of the direct precursors to the 2 series

prostaglandins.

Reproductive Measurements

Size of the previous pregnant horn at 21 and 28 DIM did not differ between

treatments or parities, nor did the change in uterine horn size from 21 to 28 DIM (Table

3.5). At 21 DIM, mulitparous cows fed CSLCFA prepartum had more uterine tonus than

multiparous cows not fed CSLCFA prepartum, however this was not evident in

primiparous animals (CSLCFA prepartum vs. no CSLCFA prepartum by parity









interaction; P = 0.050). This pattern did not carry over to measurements taken at 28

DIM. There were no differences between treatments or parities in the size of the cervical

os at 21 DIM (3.38 cm). However at 28 DIM, multiparous cows fed CSLCFA prepartum

tended to have a smaller cervical os than those not fed CSLCFA prepartum (2.97 vs. 3.30

cm) whereas this did not occur in primiparous cows fed CSLCFA prepartum versus those

not fed CSLCFA (3.12 vs. 2.89 cm) (parity by CSLCFA prepartum vs. no CSLCFA

prepartum interaction, P = 0.089). There were no differences between treatments or

parities in whether the cervix was open or closed at 21 and 28 DIM (Table 3.5). At 21

DIM, cervical color was red only for some primiparous cows fed CSLCFA prepartum.

Therefore the CSLCFA prepartum vs. no CSLCFA prepartum by parity interaction was

significant (P < 0.001). Cervical color at 28 DIM was not affected by treatment or parity

(Table 3.5).

Clinical endometritis as described by a purulent or foul discharge after 20 d

postpartum or a mucopurulent discharge after 26 d postpartum was associated with a

reduction of pregnancy rates (LeBlanc et al., 2002). In addition, abnormal vaginal

discharge has been correlated with a delay in the first postpartum ovulation (Opsomer et

al., 2000). Furthermore, if the first ovulation occurred in the presence of a uterus with

heavy contamination, it led to prolonged luteal phases which was also associated with

lower fertility (Opsomer et al., 2000). Primiparous cows had a greater (more purulent

discharge) average score for vaginal discharge amount at 21 DIM in comparison to

multiparous cows (2.32 vs. 1.77; P = 0.034), however at 28 DIM there were no

differences between treatments or parities (Table 3.6). Primiparous cows had a greater

average score for vaginal discharge quality (greater evidence of infection) at 21 DIM in









comparison to multiparous cows (3.37 vs. 1.42; P < 0.001), however at 28 DIM

differences among treatments or between parities were not detected (Table 3.6).

Primiparous cows had less class 1 (P = 0.048) and class 2 follicles (P = 0.034) at 21

DIM in comparison to multiparous cows (Table 3.7), but the number of class 3 follicles

tended to be greater at 28 DIM (P = 0.099) in primiparous compared to multiparous cows

(Table 3.7). The number of class 1 (8.0 vs. 15.5; P = 0.09) and class 2 (0.7 vs. 2.8; P =

0.10) follicles at 21 DIM was less in multiparous cows fed CSLCFA prepartum compared

to cows not fed CSLCFA prepartum (Table 3.7). However, there were no such changes

in primiparous cows thus accounting for the interaction. These changes in follicle

dynamics at 21 DIM reflects stimulation in the number of class 3 follicles for multiparous

cows due to fat feeding prepartum. Number of class 3 follicles at 21 DIM were increased

in multiparous cows fed CSLCFA prepartum (2.3 vs. 1.4; P = 0.06) with no differences

among diets for the number of class 3 follicles of primiparous cows. This early stimulus

in pre-ovulatory follicles (class 3) due to fat feeding prepartum was eliminated among the

other groups by 28 DIM with primiparous cows tending to have greater overall number of

class 3 follicles (2.2 vs. 1.6; P = 0.10). At 21 DIM, the number of CL tended to be fewer

(0.4 vs. 0.9) and smaller (16.9 vs. 25.2 mm) for cows fed CSLCFA starting at 1 DIM

compared to those not fed CSLCFA (Table 3.7). The number and size of CL present on

the ovaries at 28 DIM did not differ between treatments or parities.

Stimulation of ovarian follicle development in cows fed supplemental fat often has

been reported (Staples et al., 1998). Fat supplementation for 21 d prepartum did not

affect follicle dynamics in cows fed isoenergetic diets containing 1.7% supplemental fat

(prilled long chain FA) (Frajblat and Butler, 2003). However supplemental fat prepartum









resulted in greater pregnancy rates (86 vs. 58%). In contrast, workers in Missouri

(Oelrichs et al., 2004) recently reported that cows fed soybeans beginning either

prepartum or at calving had fewer small (< 5 mm) follicles and tended to have more

medium size (6 to 9 mm) follicles than controls during the first synchronized estrous

cycle. During the first estrous synchronization in the present study, there was no

difference in number of class 2 follicles (6 to 9 mm), class 3 follicles (> 10 mm), number

of CL, or size of CL present as determined by ultrasound on the day that GnRH was

injected (d 0), nor on d 2, 4, 6, or 7 following GnRH (Table 3.8). Such changes were

evident in the present study through 21 and 28 d postpartum. However, changes

thereafter during the synchronization period were not detected. For example, during the

same time frame, the number of class 1 follicles (2 to 5 mm) was fewer for cows fed

CSLCFA prepartum compared to cows fed CSLCFA beginning in the postpartum period

(10.4 vs. 14.8 averaged across the five measurement periods; P = 0.008). This may

reflect a greater turnover of follicles in this group.

Improvement in conception rates when fat is supplemented in the diet is reported

often (Staples et al., 1998). No change in conception rates due to fat feeding also is

regularly reported (Holter et al., 1992; Oelrichs et al., 2004). In contrast, a few studies

have reported a decrease in conception rates of cows fed supplemental fat (Erickson et

al., 1992; Sklan et al., 1994), although it may have been due to greater milk production

and a more negative energy balance which have been strongly linked to decreased

fertility in dairy cattle. In this study, first service conception rates of cows that responded

to synchronized ovulation were 27% (3/11), 40% (4/10), 70% (7/10), and 63% (7/11) for

cows fed no CSLCFA, CSLCFA prepartum, CSLCFA beginning at 1 DIM, and CSLCFA









beginning at 28 DIM, respectively. Cows fed CSLCFA tended to have greater first

service conception rates compared to cows not fed CSLCFA (58 vs. 27%, respectively, P

< 0.10).

The number of cows per treatment in this study were too low to have a lot of

confidence in the conception rate results however a 31% increase is noteworthy.

Although improvement in first service conception rate was not due to differences in

follicular dynamics during the first synchronized estrous, IGF-I acts synergistically with

LH to promote follicular development (Lucy, 2001). Greater concentrations of plasma

IGF-I around the time of breeding in multiparous cows fed CSLCFA (Figure 3.9) may

have contributed to increased first service conception rates.

Hepatic Measurements

In the present study, mean hepatic TAG concentrations tended to be greater for

cows fed CSLCFA beginning prepartum in comparison to cows not fed CSLCFA

prepartum (16.7 vs. 10.4% of dry liver weight; P = 0.080; Table 3.9). Hepatic TAG

concentrations in cows fed CSLCFA prepartum and control diets peaked at 14 DIM,

however, cows fed CSLCFA beginning at 1 DIM did not decrease at 28 DIM but

remained elevated (treatment by day interaction; P = 0.04; Figure 3.15). At 14 DIM the

TAG concentration of cows fed CSLCFA prepartum tended to be greater than that of

cows not fed CSLCFA prepartum (P = 0.084; Figure 3.15). Plasma NEFA

concentrations were greater in cows fed CSLCFA prepartum than cows not fed CSLCFA

prepartum in early lactation, which may account for the differences in TAG

accumulation. In contrast, Illinois workers (Grum et al., 1996) reported decreased liver

TAG concentrations at 1 DIM and a tendency for decreased plasma NEFA concentrations

early postpartum in cows fed 6.5% of DM as fat for 50 d prepartum. They also reported a









positive correlation between concentrations of plasma NEFA at 3 d prepartum and the

concentration of TG in the liver at 1 d postpartum. However, a later study (Douglas et

al., 2004) in which cows were fed a moderate non-fiber carbohydrate (NFC) control diet,

a low NFC diet supplemented with 4% fat prepartum and 2% fat postpartum, or a

moderate NFC fat-supplemented diet beginning at 60 d prepartum revealed no treatment

differences in DMI, milk production, plasma concentration of NEFA, or total hepatic

lipid or TAG at 1 DIM. Additionally, recent research conducted at the University of

Florida found no difference in hepatic TAG accumulation in cows fed control, 225 g/d of

Ca salts of conjugated linoleic acid (CLA), or 225 g/d of Ca salts of trans C18:1

beginning at 28 d prepartum despite differences in plasma NEFA concentrations at 1 wk

postpartum (Selberg et al., 2004). While others have reported a negative effect of

elevated hepatic TAG on day to first estrus (Jorritsma et al., 2000) or to first ovulation

(Marr et al., 2002), days to first ovulation in this study was not greater for the treatment

group with elevated hepatic TAG concentration (CSLCFA prepartum).

Neither treatment nor parity affected hepatic IGF-I or IGF-II mRNA expression

(Table 3.9; Figures 3.17 and 3.18). The liver is the primary source of circulating IGF-I

and decreased hepatic IGF-I mRNA expression results in decreased circulating

concentrations of IGF-I in cattle (Wang et al., 2003). In this study, the additional hepatic

lipid present postpartum in cows fed CSLCFA prepartum was not detrimental to IGF-I or

IGF-II mRNA expression. Cows fed CSLCFA beginning prepartum tended to have less

IGFBP-2 mRNA expression in comparison to cows not fed CSLCFA prepartum (0.078

vs. 0.121 arbitrary units/18S; P = 0.097; Figure 3.19). Steady state expression of IGF-II

mRNA expression, which is an indirect measurement of synthesis, was not affected by









treatment and steady state IGFBP-2 expression was lower in cows fed CSLCFA

prepartum, indirectly implying that circulating IGF-II in cows fed CSLCFA prepartum

was increased. Recently, University of Florida workers reported no treatment differences

in hepatic IGF-I mRNA expression in cows fed a control diet, 225 g/d of calcium salts of

CLA, or 225 g/d of trans C18:1 during early lactation (Selberg et al., 2003). However,

IGF-II mRNA and IGFBP-2 abundance were greater for cows fed trans C18:1 than for

cows fed the control diet or CLA. Expression of mRNA of IGF-1 (Figure 3.17), IGF-II

(Figure 3.18), and IGFBP-2 (Figure 3.19) did not differ among biopsy days nor were

treatment by day interactions detected to be significant for these three dependent

variables.

Measurements of Immune Status

Immune reactions have been shown to be modulated by the diet, including the

PUFA composition of the diet (Calder et al., 2002). Mechanisms involved in regulation

are not yet understood, but evidence exists that the poly unsaturated FA composition of

the diet influences cellular communication and activation through the synthesis of

prostaglandins, tumor necrosis factor-a, and interferon-y (Calder et al., 2002). Linoleic

acid can be converted to arachidonic acid, the precursor for prostaglandin E2 and

leukotriene B4 which are pro-inflammatory mediators. Lessard et al. (2004) evaluated

cellular immune functions of dairy cows fed supplemental fat during the transition

period. Cows were fed a diet containing 2.7% Ca salt of palm oil, 5.9% flaxseed (n-3

FA), or 9.4% micronized soybeans (n-6 FA) from 6 wk prepartum to calving followed by

diets containing and 4.7% Ca salt of palm oil, 9.7% flaxseed, or 20.3% micronized

soybeans from calving to 6 wk postpartum. The authors concluded that cellular immune

functions were modulated around parturition; however feeding diets rich in n-3 or n-6 FA









did not have a major impact on cellular immune function. In the present study, incidence

of disease mastitiss, metritis, or RFM) during the first 10 DIM was lower (P < 0.05) for

cows fed CSLCFA prepartum (8%, 1/12) versus those not fed CSLCFA prepartum (35%

[8/23] for the no CSLCFA treatment and 58% [7/12] for the CSLCFA at 1 DIM

treatment) which could have had beneficial effects on milk production and reproduction.

In response to immunological stress, the liver will produce the acute phase proteins

ceruloplasmin, fibrinogen, and haptoglobin (Baumann and Gauldie, 1994). It is unclear

whether increased concentrations of acute phase proteins in lactating dairy cattle is due to

greater stress (indicating an adverse state) or due to a greater immune response

(indicating a healthier state). Fibrinogen is involved with blood clotting and the

formation of the fibrin matrix for tissue repair. Increased fibrinogen concentrations are

detected during internal hemorrhage or tissue damage. Normal values in cattle range

from 100 to 600 mg/dl (The Merck Veterinary Manual, 1997). In this study, the range

was 26 to 402 mg/dl. The concentrations rose, plateaued, then decreased during the first

4 wk postpartum. Mean plasma concentrations of fibrinogen during the first 27 d

postpartum tended to be greater for control cows than for cows fed CSLCFA beginning at

1 DIM (123.8 vs. 100.2 mg/dl; P = 0.07; Table 3.10). Primiparous cows had greater

fibrinogen concentrations when CSLCFA was withheld prepartum than when fed

prepartum (114.4 vs. 70.5 mg/dl); however fibrinogen concentrations were not different

for multiparous cows regardless of when CSLCFA feeding was initiated (no CSLCFA

prepartum vs. CSLCFA prepartum by parity interaction; P = 0.029; Table 3.10).

Haptoglobin is responsible for binding iron, a limiting nutrient in bacterial growth.

Because haptoglobin concentrations are normally undetectable in bovine blood unless









there is tissue damage, it can be a good indicator of the immunological stress response to

parturition. In this study, the range was 0 to 329 mg ofHbB/ 100 ml (the amount of

hemoglobin bound by haptoglobin/ 100 ml of plasma). Mean plasma concentrations of

haptoglobin during the first 27 d postpartum were greater for primiparous than

multiparous cows (31.9 vs. 17.0 mg of HbB/ 100 ml of plasma; P = 0.043; Table 3.10),

indicating that parturition caused a greater immune response in primiparous cows. Using

heterogeneity of regression, the cubic pattern of plasma haptoglobin over time was

affected by feeding CSLCFA and parity (Table A-5). Primiparous cows not fed

CSLCFA prepartum had greater initial plasma concentrations of haptoglobin than cows

fed CSLCFA prepartum and declined at a faster rate so that both treatment groups

reached their nadir at 23 DIM (Figure 3.20A). Multiparous cows fed CSLCFA

prepartum had very low initial concentrations of haptoglobin, rose to peak concentrations

at 7 DIM and reached their nadir at 23 DIM. Multiparous cows not fed CSLCFA

prepartum had higher initial concentrations and steadily declined through 27 DIM (Figure

3.20B). The trends over time indicate cows fed CSLCFA prepartum had less of an acute

phase protein response to parturition than cows not fed CSLCFA prepartum, although it

is unclear why haptoglobin concentrations in multiparous cows fed CSLCFA prepartum

did not peak until 7 DIM.

Ceruloplasmin is involved with copper transport to tissues utilizing its antioxidant

properties and concentrations will increase due to an inflammatory response of the cow.

Normal values in cattle range from 16.8 to 34.2 mg/dl (The Merck Veterinary Manual,

1997). In this study, the range was from 9.0 to 51.0 mg/dl, suggestive of an acute

inflammatory response to parturition. Mean plasma concentrations of ceruloplasmin









were greater for primiparous than for multiparous cows during the first 27 d postpartum

(24.4 vs. 20.7 mg/dl; P = 0.007) indicating that parturition caused a greater immune

response in primiparous cows. The pattern over time was not different among treatments

or between parities (Figure 3.21).

Elevations in plasma concentrations of total biliruben are due to severe hepatic

damage or extrahepatic obstruction, and following pregnancy. Normal values in cattle

range from 0 to 0.5 mg/dl (The Merck Veterinary Manual, 1997). In this study, the range

was 0.0 to 1.3 mg/dl. Mean plasma concentrations of total biliruben were greater for

cows fed CSLCFA prepartum than for cows not fed CSLCFA prepartum (0.36 vs. 0.23

mg/dl; P = 0.04), although values were still within the normal range. An effect of week

postpartum was detected (P < 0.001; Figure 3.22).

Elevations in plasma concentrations of ALT are due to hepatic disease although it

is not a good indicator in cattle because of its low activity and it is not liver specific (The

Merck Veterinary Manual, 1997). Mean plasma concentrations of ALT did not differ

among treatments or between parities (21.6 IU/L; Table 3.10), however an effect of week

postpartum was detected (P = 0.003; Figure 3.23).

Elevations in plasma concentrations of ALK are due to liver lesions or bile duct

obstruction, however in cattle there is a very wide range of normal activity between

animals (The Merck Veterinary Manual, 1997). Mean plasma concentrations of ALK

were lower for multiparous cows in comparison to primiparous cows (31.7 vs. 44.1 U/L;

Table 3.10). An effect of week postpartum was detected (P < 0.001; Figure 3.24).

Concentrations of AST are present in most tissues and it is not liver specific.

Damage to the liver and the postparturient period in cattle cause leakage of large amounts









of AST into blood (The Merck Veterinary Manual, 1997). Mean plasma concentrations

of AST did not differ among treatments or between parities (90.5 IU/L; Table 3.10),

however an effect of week postpartum was detected (P = 0.01; Figure 3.25).

Plasma concentrations of GGT are of hepatic origin and can be a good indicator of

liver and bile duct damage in cattle (The Merck Veterinary Manual, 1997). Mean plasma

concentrations of GGT did not differ among treatments or between parities (30.0 U/L;

Table 3.10), however an effect of week postpartum was detected (P = 0.01; Figure 3.25).

At wk 4 postpartum cows fed CSLCFA prepartum had greater plasma concentrations of

GGT than cows not fed CSLCFA prepartum (P = 0.03; Figure 3.26).

Plasma concentrations of albumin decrease due to the failure of hepatic

parenchymal synthesis. In cattle, only 5% of serum albumin levels are synthesized per

day, therefore it takes time to see hepatic damage (The Merck Veterinary Manual, 1997).

Concentration of plasma albumin increased slightly with wk postpartum (P = 0.003;

Figure 3.27). Initiation of CSLCFA supplementation at calving stimulated albumin

concentrations in plasma of primiparous cows but reduced it in multiparous cows (no

CSLCFA vs. CSLCFA 1 DIM by parity interaction; P = 0.018; Table 3.10).

Parity Effects

As expected, primiparous cows weighed less, produced less milk, and tended to

have less SCC in the milk than multiparous cows. In addition, primiparous cows had

greater circulating glucose and tended to have lower circulating BUN then multiparous

cows. Parturition, removal of the calf, and the milking process is probably more stressful

on first calf heifers than on multiparous cows. Indicative of a higher stress level, heifers

had a greater immune response soon after parturition than cows as indicated by greater

circulating concentration of the acute phase proteins, haptoglobin and ceruloplasmin, and









the liver enzyme ALK. At 21 DIM, primiparous cows had more vaginal discharge, a

more purulent vaginal discharge, and fewer class 1 and class 2 follicles and tended to

have more class 3 follicles at 28 DIM than multiparous cows.

Conclusion

Holstein cows began consuming a diet at 0 or 2% of dietary DM as CSLCFA at

-28 d prepartum. Cows fed the 0% CSLCFA diet either remained on a CSLCFA-free

diet at parturition or were shifted to a 2% CSLCFA diet at either 1 or 28 DIM and

remained on said diet until 100 DIM. Animals fed CSLCFA in the prepartum period

continued to receive CSLCFA throughout the lactation period. Cows fed CSLCFA

beginning in the perpartum period tended to produce more milk. This milk increase was

accompanied by a tendency for elevated concentrations of TG in the liver at 14 DIM, a

tendency for lower expression of hepatic IGFBP-2 mRNA, and elevated concentrations

of plasma biliruben compared to cows not receiving CSLCFA prepartum. Multiparous

cows appeared to benefit more from supplementation with CSLCFA than primiparous

cows in that multiparous cows not fed CSLCFA at any time during the study had or

tended to have lower concentrations of milk protein, longer and greater loss of BW,

greater concentrations of plasma BHBA, and lower concentrations of plasma glucose,

leptin, and IGF-I. In addition, multiparous cows fed CSLCFA prepartum tended to have

fewer small and medium size but more larger size follicles (> 10 mm in size), more

uterine tone at 21 DIM a smaller cervical os at 28 DIM, and a slower decrease in plasma

concentrations of PGFM the first 10 d postpartum than multiparous cows not fed

CSLCFA prepartum. Multiparous cows fed CSLCFA had greater concentrations of IGF-

I at the time of AI and all cows fed CSLCFA tended to have greater conception rate at

first service regardless of day of initiation of CSLCFA supplementation.









Table 3.1: Ingredient and chemical composition of diets fed to nonlactating cows.
Control CSLCFA1
Ingredient, % of DM
Corn silage 50 50
Bermudagrass hay 10 10
Ground corn 16.9 14.4
Citrus pulp 5.0 5.1
Soybean meal 11.8 12.4
Megalac- R1 0.0 2.0
Mineral mix 2 6.2 6.2
Trace mineral salt 3 0.009 0.009

Chemical
DM% 39.8 39.8
NEL, Mcal/kg of DM 1.58 1.61
CP, % of DM 15.1 14.3
Ether extract, % of DM 3.1 4.4
NDF, % of DM 36.2 36.7
ADF, % of DM 21.3 21.6
Ca, % of DM 2.2 2.1
P, % of DM 0.33 0.32
K, % of DM 1.1 1.1
Mg, % of DM 0.35 0.33
Na, % of DM 0.24 0.25
Cl, % of DM 1.04 0.92
S, % of DM 0.26 0.26
Mn, mg/kg of DM 34 33
Cu, mg/kg of DM 12 14
Zn, mg/kg of DM 42 47
Fe, mg/kg of DM 257 249
Mo, mg/kg of DM 1.4 1.6
1Megalac- R, Church & Dwight Co. Inc., Princeton, NJ.
2 Mineral Mix contained 22.8% CP, 22.9% Ca, 0.20% P, 0.2% K, 2.8% Mg, 0.7% Na,
2.4% S, 8% Cl, 147 mg/kg of Mn, 27 mg/kg of Fe, 112 mg/kg of Cu, 95 mg/kg of Zn, 7
mg/kg of Se, 8 mg/kg of I, 11 mg/kg of Co, 268,130 IU of vitamin A/kg, 40,000 IU of
vitamin D/kg, and 1129 IU of vitamin E/kg (DM basis).
3 Minimum concentrations of 40% Na, 55% Cl, 0.25% Mn, 0.2% Fe, 0.033% Cu,
0.007% I, 0.005% Zn, and 0.0025% Co (DM basis).









Table 3.2: Ingredient and chemical composition of diets fed to lactating cows.
Control CSLCFA1
Ingredient, % of DM
Corn silage 37.5 37.5
Alfalfa hay 10 10
Ground corn 24.2 21.9
Citrus pulp 5.0 5.0
Cottonseed hulls 2.5 2.5
Soybean meal 9.1 9.4
Soy Plus 2 6.8 6.8
Bio Phos 3 0.4 0.4
Megalac- R1 0.0 2.0
Mineral mix 4 4.5 4.5

Chemical
DM% 46.2 46.3
NEL, Mcal/kg of DM 1.67 1.71
CP, % of DM 16.9 16.7
Ether extract, % of DM 3.1 4.7
NDF, % of DM 30.9 30.2
ADF, % of DM 18.9 18.8
Ca, % of DM 1.15 1.51
P, % of DM 0.49 0.46
K, % of DM 1.27 1.27
Mg, % of DM 0.29 0.31
Na, % of DM 0.44 0.53
Cl, % of DM 0.34 0.39
S, % of DM 0.23 0.24
Mn, mg/kg of DM 71 86
Cu, mg/kg of DM 23 23
Zn, mg/kg of DM 88 106
Fe, mg/kg of DM 236 227
Mo, mg/kg of DM 2.1 1.6
Megalac- R, Church & Dwight Co. Inc., Princeton, NJ.
2 West Central Soy, Ralston, IA.
3 IMC-Agrico, Bannockburn, IL.
4 Mineral Mix contained 26.4% CP, 10.2% Ca, 0.90% P, 3.1% Mg, 1.5 % S, 5.1% K,,
8.6 % Na, 11698 mg/kg of Zn, 512 mg/kg of Cu, 339 mg/kg of Fe,2231 mg/kg of Mn, 31
mg/kg of Co, 26 mg/kg of I, 7.9 mg/kg of Se, 147,756 IU of vitamin A/kg, and 787 IU of
vitamin E/kg (DM basis).











Table 3.3. Milk yield, milk composition, postpartum body weight, and postpartum body condition score of Holstein cows fed diets
without calcium salts of long chain fatty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum, at 1
day in milk (DIM) or at 28 DIM.
Treatments Orthogonal contrasts', P =
No CSLCFA CSLCFA CSLCFA
Measure CSLCFA prepartum 1 DIM 28 DIM SE A B C Parity D E F
P2 M3 P M P M P M


Milk yield,
kg/d
3.5% FCM,
kg/d
Milk fat, %
Milk protein,
%
Milk fat yield,
kg/d
Milk protein
yield, kg/d
Milk SCC,
x 1000/ml
BW, kg
BCS


.0 41.0

.1 39.7

15 3.28

53 2.47

27 1.36

)3 1.06

7 750

0 644


39

40

3.

2.

1.

1.(

46

53


.5 44.8

.2 42.5

55 3.33

58 2.69

43 1.44

06 1.25

3 466

.9 621


1 38.3

7 37.3

3 3.41

S 2.70

1 1.28


34.7

35.4

3.57

2.75

1.26

1.01

282

533


39.9

38.5

3.42

2.71

1.34

1.10

428

664


0.924

0.947

0.487

0.102

0.912

0.756

0.324

0.991


0.059

0.087

0.789

0.525

0.114

0.080

0.760

0.676


0.896

0.779

0.871

0.241

0.659

0.439

0.465

0.555


0.041

0.215

0.195

0.846

0.451

0.067

0.067

<0.001


0.933

0.861

0.977

0.091

0.865

0.487

0.614

0.992


0.816

0.920

0.782

0.679

0.783

0.649

0.400

0.325


0.710

0.941

0.949

0.261

0.967

0.797

0.407

0.957


3.06 2.77 2.88 3.10 3.02 3.31 3.13 2.95 0.19 0.363 0.501 0.534 0.937 0.220 0.626 0.231


SOrthogonal contrast of means were the following: A = No CSLCFA vs. CSLCFA, B = CSLCFA prepartum vs. (CSLCFA 1 DIM plus CSLCFA 28 DIM), C
CSLCFA 1 DIM vs. CSLCFA 28 DIM, D = contrast A by parity, E = contrast B by parity, and F = contrast C by parity.
2Primiparous cows.
3Multiparous cows.










Table 3.4. Concentration of plasma hormones and metabolites and day of first ovulation of Holstein cows fed diets without calcium
salts of long chain fatty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum, at 1 day in milk (DIM)
or at 28 DIM.
Treatments Orthogonal contrasts', P =
No CSLCFA CSLCFA CSLCFA
Measure CSLCFA prepartum 1 DIM 28 DIM SE A B C Parity D E F
P2 M3 P M P M P M
BHBA, mg/dl 10.7 14.2 10.4 9.7 10.2 6.7 8.4 6.9 1.6 0.007 0.146 0.629 0.635 0.044 0.501 0.520
NEFA, meq/L 384 450 432 480 400 241 289 245 56 0.149 0.002 0.348 0.588 0.217 0.130 0.316
Glucose, mg/dl 65.5 55.2 65.2 60.4 68.7 64.8 67.8 66.5 2.1 0.004 0.025 0.846 0.001 0.054 0.553 0.533
Blood urea
10.2 11.5 10.4 15.3 9.8 10.9 10.6 11.0 1.3 0.682 0.053 0.744 0.055 0.706 0.081 0.808
nitrogen, mg/dl
Leptin, ng/ml 3.06 1.65 2.83 2.62 1.91 1.99 1.70 2.57 0.39 0.846 0.156 0.729 0.669 0.078 0.465 0.470

IGF-I, ng/ml 77.1 54.3 62.2 60.6 64.3 69.9 66.2 74.1 7.5 0.953 0.261 0.683 0.641 0.040 0.487 0.885

Insulin, ng/ml 0.62 0.54 0.65 0.52 0.71 0.75 0.74 0.63 0.09 0.236 0.102 0.608 0.320 0.975 0.536 0.449
Accumulated
progesterone, 22.0 27.4 24.0 24.0 21.9 26.2 26.5 23.3 4.0 0.948 0.884 0.779 0.555 0.516 0.906 0.602
1 to 77 DIM,
ng/ml
Accumulated
progesterone, 7.3 11.1 8.7 10.9 7.9 9.7 9.1 8.0 2.4 0.942 0.806 0.930 0.467 0.606 0.931 0.847
1 to 46 DIM,
ng/ml
DIM at first
27.8 26.3 29.7 27.8 32.2 27.5 22.8 24.6 4.0 0.910 0.560 0.125 0.587 0.987 0.955 0.417
ovulation
1 Orthogonal contrast of means were the following: A = No CSLCFA vs. CSLCFA, B = CSLCFA prepartum vs. (CSLCFA 1 DIM plus CSLCFA 28 DIM), C =
CSLCFA 1 DIM vs. CSLCFA 28 DIM, D = contrast A by parity, E = contrast B by parity, and F = contrast C by parity.
2Primiparous cows.
3 Multiparous cows.











Table 3.5. Concentration of plasma PGF2a metabolite (PGFM) the first 14 DIM and the size and characteristics of the uterus and
cervix of Holstein cows fed diets without calcium salts of long chain fatty acids (CSLCFA) (no CSLCFA) or diets with
CSLCFA starting prepartum, or at 1 day in milk (DIM).
Treatments Orthogonal contrasts', P =
No CSLCFA CSLCFA


Measure CSLCFA


PGFM, pg/ml

Previous pregnant horn size
at 21 DIM, cm
Previous pregnant horn size
at 28 DIM, cm
Change in uterine horn size
from 21 to 28 DIM, cm
Uterine tonus4 at 21 DIM
Uterine tonus4 at 28 DIM
Size of cervical os at 21
DIM, cm
Size of cervical os at 28
DIM, cm
Cervix closed (1) or open
(2) at 21 DIM
Cervix closed (1) or open
(2) at 28 DIM
Cervical color, pink (1) or
red (2) at 21 DIM
Cervical color, pink (1) or
red (2) at 28 DIM


P2 M3

1004 1186

2.43 2.52

2.24 2.60

-0.20 0.08

1.80 1.77
1.70 1.69

3.20 3.59

3.12 3.23

1.20 1.00

1.10 1.00

1.00 1.00


prepartum 1 DIM SE
P M P M

1443 1412 1536 955 244

2.38 2.62 2.55 3.95 0.18

2.50 2.35 2.42 2.59 0.20

0.12 -0.27 -0.13 -0.36 0.20

1.50 2.17 2.17 1.50 0.28
1.33 2.00 1.67 1.83 0.26

3.35 3.54 3.25 3.38 0.19

3.12 2.97 2.67 3.38 0.18

1.00 1.00 1.00 1.00 0.07

1.00 1.00 1.00 1.00 0.05

1.50 1.00 1.00 1.00 0.07


1.00 1.08 1.17 1.00 1.00 1.00 0.08


A B Parity C D


0.261

0.500

0.841

0.683

0.923
0.815

0.610

0.727

0.460

0.622

< 0.001


0.722 0.418

0.133 0.114

0.655 0.452

0.357 0.456

0.855 0.964
0.832 0.209

0.663 0.140

0.387 0.136

0.164 0.277

0.350 0.467

1.00 0.007


0.367 0.606 0.639


0.509 0.111

0.984 0.376

0.256 0.613

0.269 0.199

0.050 0.237
0.227 0.731

0.852 0.480

0.089 0.086

0.460 0.164

0.622 0.350

< 0.001 1.00

0.152 0.606


1 Orthogonal contrast of means were the following: A = (No CSLCFA plus CSLCFA 1 DIM) vs. CSLCFA prepartum, B = No CSLCFA vs. CSLCFA 1 DIM,
C = contrast A by parity, and D = contrast B by parity.
SPrimiparous cows.
3Multiparous cows.
4 Score of 1 = no tonus, 2 = moderate tonus, and 3 = tonus.










Table 3.6. Vaginal observations at 21 and 28 DIM of Holstein cows fed diets without calcium salts of long chain fatty acids
(CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum, or at 1 day in milk (DIM).
Treatments Orthogonal contrasts', P =
No CSLCFA CSLCFA
Measure CSLCFA prepartum 1 DIM SE A B Parity C D
P2 M3 P M P M

Vaginal discharge
Vaginal discharge 2.30 2.15 2.33 1.50 2.33 1.67 0.30 0.481 0.441 0.034 0.445 0.377
amount4 at 21 DIM
Vaginal discharge
Vaginal discharge 2.40 1.46 1.83 2.00 2.17 1.83 0.33 0.872 0.828 0.182 0.189 0.344
amount4 at 28 DIM
Vaginal discharge 3.60 1.92 2.67 1.00 3.83 1.33 0.60 0.133 0.759 <0.001 0.702 0.479
quality5 at 21 DIM
Vaginal discharge 2.50 1.08 1.50 167 3.33 2.00 0.69 0313 0.193 0.137 0.228 0.946
quality5 at 28 DIM
SOrthogonal contrast of means were the following: A = (No CSLCFA plus CSLCFA 1 DIM) vs. CSLCFA prepartum, B = No CSLCFA vs. CSLCFA 1 DIM,
C = contrast A by parity, and D = contrast B by parity.
2Primiparous cows.
3Multiparous cows. oo
4 Score of 1 = none, 2 = trace, 3 = slight, 4 = moderate, or 5 = copious.
5 Score of 0 = none, 1 = clear, 2 = cloudy, 3 = mucous with pus, 4 = mucopurulent, or 5 = purulent.











Table 3.7. The number and size1 of ovarian structures at 21 and 28 DIM of Holstein cows fed diets without calcium salts of long chain
fatty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum, or at 1 day in milk (DIM).
Treatments Orthogonal contrasts2, P =


No
CSLCFA


CSLCFA
prepartum


CSLCFA
1 DIM


SE A B Parity


Number of class 1 follicles
at 21 DIM
Number of class 2 follicles
at 21 DIM
Number of class 3 follicles
at 21 DIM
Number of CL5 at 21 DIM

Size of CL at 21 DIM, mm
Number of class 1 follicles
at 28 DIM
Number of class 2 follicles
at 28 DIM
Number of class 3 follicles
at 28 DIM
Number of CL at 28 DIM
Size of CL at 28 DIM, mm


P3 M4 P M P M

8.4 12.8 9.0 8.0 10.7 18.3 2.2 0.050 0.074

1.1 2.8 0.8 0.7 0.5 2.8 0.7 0.111 0.695

1.4 1.5 1.0 2.3 1.8 1.3 0.4 0.718 0.780

0.7 1.2 0.8 1.0 0.3 0.5 0.3 0.442 0.080

22.9 27.5 23.0 29.0 12.3 21.5 4.6 0.256 0.084

14.0 14.3 10.5 9.7 12.0 16.3 2.3 0.058 0.995

2.0 2.5 1.3 1.5 0.7 1.3 0.7 0.723 0.055

2.1 1.7 2.7 1.8 1.8 1.5 0.4 0.181 0.530

0.9 0.9 0.7 1.2 0.8 1.2 0.3 0.880 0.746
27.1 24.3 27.0 34.4 33.2 26.9 5.3 0.581 0.396


0.048

0.034

0.358

0.278

0.103

0.504

0.411

0.099

0.227
0.903


0.087

0.100

0.057

0.756

0.915

0.454

0.723

0.505

0.536
0.295


0.451

0.627

0.437

0.555

0.616

0.365

0.921

0.919

0.571
0.729


1 Class 1 follicles were 2 to 5 mm, class 2 follicles were 6 to 9 mm, and class 3 follicles were > 10 mm.
2 Orthogonal contrast of means were the following: A = (No CSLCFA plus CSLCFA 1 DIM) vs. CSLCFA prepartum, B = No CSLCFA vs. CSLCFA 1 DIM,
C = contrast A by parity, and D = contrast B by parity.
3Primiparous cows.
4Multiparous cows.
5 Corpus luteum.


Measure











Table 3.8.Ovarian structures present on the ovaries of Holstein cows fed diets without calcium salts of long chain fatty acids
(CSLCFA) (no CSLCFA) or diets with CSLCFA starting prepartum, at 1 day in milk (DIM) or at 28 DIM where day
equals the days after GnRH injection for estrous synchronization (44 + 3 DIM).
Treatments


Measure
Number of class 1 follicles a
(2 to 5 mm)





Number of class 2 follicles
(6 to 9 mm)




Number of class 3 follicles
(> 10 mm)




Number of corpus luteum b


S


No
CSLCFA
14.0
11.2
12.6
12.3
12.8

1.5
2.2
2.3
2.4
1.8
2.3
2.1
2.2
2.4
2.8
1.0
1.0
1.2
1.3
1.4


ize of corpus luteum 0 21.3
(mm) 2 21.3
4 25.8
6 27.3
7 28.7
a CSLCFA prepartum vs. CSLCFA postpartum, P =
b CSLCFA 1 DIM vs. CSLCFA 28 DIM, P = 0.099.
CSLCFA 1 DIM vs. CSLCFA 28 DIM, P = 0.092.


CSLCFA
prepartum
9.1
11.5
9.9
11.0
10.5

1.0
3
1.4
2.3
1.9
1.8
1
2.1
2.3
2.5
0.7
0.9
1.25
1.3
1.5
11.6
16.7
21.0
25.3
29.0
0.008.


CSLCFA
1 DIM
15.2
15.3
16.3
15.7
16.4

3.25
3.4
1.4
0.9
2
2.5
1.3
2.3
1.6
2.0
0.7
0.9
1.2
1.0
1.2
13.4
15.4
21.7
20.2
25.5


CSLCFA
28 DIM
13.4
19.1
13.8
12.1
10.3

2.1
2.1
3.2
2.4
1.6
2.0
1.1
1.9
1.9
2.0
1.0
1.2
1.3
1.7
1.7
21.8
25.0
26.6
34.6
29.6










Table 3.9. Concentrations of hepatic triacylglycerol (TAG), and hepatic IGF-I, IGF-II, and IGF binding protein (BP) -2 mRNA levels
of Holstein cows fed diets without calcium salts of long chain fatty acids (CSLCFA) (no CSLCFA) or diets with CSLCFA
starting prepartum, or at 1 day in milk (DIM).


Treatments Orthogonal contrasts', P =

No CSLCFA CSLCFA
Measure CSLCFA prepartum 1 DIM SE A B Parity C D
P2 M3 P M P M
TAG, % of wet
2.8 3.0 5.3 6.0 2.7 5.6 1.3 0.102 0.317 0.257 0.725 0.266
weight
TAG,% of dry 8.6 9.8 16.2 17.2 8.3 14.7 3.8 0.080 0.531 0.364 0.693 0.478
weight
IGF-I mRNA,
IGF-I mRNA, 0.024 0.020 0.013 0.003 0.011 0.048 0.073 0.110 0.550 0.494 0.231 0.110
arbitrary units
IGF-II mRNA,
IGF-11 mRNA, 0.292 0.288 0.269 0.224 0.296 0.331 0.043 0.159 0.591 0.892 0.429 0.670
arbitrary units
IGFBP-2 mRNA,
GFBP-2 mRNA, 0.087 0.136 0.091 0.065 0.149 0.111 0.028 0.097 0.534 0.837 0.519 0.142
arbitrary units


SOrthogonal contrast of means were the following: A = (No CSLCFA plus CSLCFA 1 DIM) vs. CSLCFA prepartum, B = No CSLCFA vs. CSLCFA 1 DIM,
C = contrast A by parity, and D = contrast B by parity.
2Primiparous cows.
3 Multiparous cows.











Table 3.10. Concentration of plasma acute phase proteins and liver enzymes of Holstein cows fed diets without calcium salts of long


chain fatty acids (CSLCFA) (no CSLCFA) or diets with
Treatments


CSLCFA starting prepartum, or at 1 day in milk (DIM).
Orthogonal contrasts P =


No
CSLCFA


Measure


Fibrinogen, mg/dl
Haptoglobin,
mg HbB/100 ml4
Ceruloplasmin, mg/dl

Total biliruben, mg/dl
Alanine
aminotransferase,
IU/L
Alkaline phosphatase,
U/L
Aspartate
aminotransferase,
IU/L
Gamma glutamyl
transferase, U/L
Albumin, g/dl


CSLCFA
prepartum


CSLCFA
1 DIM


P2 M3 P M P M
131.7 115.8 70.5 121.6 97.1 103.2

31.6 15.4 27.5 18.2 36.5 17.5

24.4 21.1 25.1 19.0 23.7 21.9

0.22 0.26 0.32 0.39 0.23 0.22

22.5 20.2 24.7 20.7 22.7 18.5


39.8


32.4 45.8 33.8 46.7 29.0


111.9 89.6 106.9 114.8


119.6 82.5


30.8 25.4 32.8 33.4 26.5 31.0

2.55 2.73 3.01 2.78 2.95 2.45


1 Orthogonal contrast of means were the following: A = (No CSLCFA plus CSLCFA
C = contrast A by parity, and D = contrast B by parity.
2Primiparous cows.
3 Multiparous cows.
4 Amount of hemoglobin bound by haptoglobin/100 ml of plasma.


SE A B Parity


13.4 0.175 0.070 0.211


C D


0.029 0.353


8.4 0.750 0.980 0.043 0.646 0.872

1.6 0.634 0.974 0.007 0.195 0.675

0.06 0.040 0.837 0.529 0.658 0.640

2.7 0.482 0.7749 0.122 0.883 0.719


3.9 0.431 0.646 < 0.001 0.947 0.172


19.3 0.576 0.986 0.289 0.294 0.692


4.5 0.264 0.880 0.976 0.900 0.265


0.15 0.104


1 DIM) vs. CSL


0.672 0.130 0.772 0.018
,CFA prepartum, B = No CSLCFA vs. CSLCFA 1 DIM,